Histone deacetylase inhibitors and compositions and methods of use thereof

ABSTRACT

Provided are certain histone deacetylase (HDAC) inhibitors of Formula I, compositions thereof, and methods of their use.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit under 35 U.S.C. § 119(e) of U.S.Provisional Application 62/158,363, filed on May 7, 2015, which ishereby incorporated by reference in its entirety.

FIELD

Provided herein are certain histone deacetylase (HDAC) inhibitors,compositions thereof, and methods of their use.

BACKGROUND

Histone deacetylases (HDACs) are zinc-containing enzymes which catalyzethe removal of acetyl groups from the ε-amino termini of lysine residuesclustered near the amino terminus of nucleosomal histones. There are 11known metal-dependent human histone deacetylases, grouped into fourclasses based on the structure of their accessory domains. Class Iincludes HDAC1, HDAC2, HDAC3, and HDAC8 and have homology to yeast RPD3.HDAC4, HDAC5, HDAC7, and HDAC9 belong to Class IIa and have homology toyeast HDAC1. HDAC6 and HDAC10 contain two catalytic sites and areclassified as Class IIb, whereas HDAC11 has conserved residues in itscatalytic center that are shared by both Class I and Class IIdeacetylases and is sometimes placed in Class IV.

SUMMARY

Provided is a compound of Formula I:

or a pharmaceutically acceptable salt, an optical isomer, or a mixtureof optical isomers thereof;

wherein:

-   -   W is N or CR⁵; X is N or CR⁶; Y is N or CR⁷; and Z is N or CR⁸;        provided not more than two of W, X, Y, and Z are N;    -   R¹ is selected from H and C₁-C₃ alkyl;    -   R² is C₂-C₃ alkylene optionally substituted with C₁-C₂ haloalkyl        or 3 or 4-membered cycloalkyl;    -   R³ and R⁴, together with the nitrogen atom to which they are        attached, form:        -   a 4, 5, 6, or 7-membered heteromonocyclic group, or        -   a 6, 7, 8, 9, or 10-membered heterobicyclic group,        -   each of which is optionally substituted with one to five            substituents each independently selected from: C₁-C₃ alkoxy,            C₁-C₃ alkyl, C₁-C₃ haloalkoxy, C₁-C₃ haloalkyl, 3 or            4-membered cycloalkoxy, 3 or 4-membered cycloalkyl, 3 or            4-membered heterocycloalkyl, carboxy, aryl, cyano, halo, and            heteroaryl, wherein aryl and heteroaryl are optionally            further substituted with one to five substituents each            independently selected from C₁-C₃ alkyl, C₁-C₃ haloalkyl,            and halo,    -   provided that if R³ and R⁴, together with the nitrogen atom to        which they are attached,    -   form a 5 or 6-membered heteromonocyclic group, then:        -   i) R² is C₂-C₃ alkylene substituted with C₁-C₂ haloalkyl or            3 or 4-membered cycloalkyl, or        -   ii) R⁸ is halo or C₁-C₃ alkyl, or        -   iii) the 5 or 6-membered heteromonocyclic group is            substituted with one to five substituents each independently            selected from: C₁-C₃ alkoxy, C₁-C₃ alkyl, C₁-C₃ haloalkoxy,            C₁-C₃ haloalkyl, 3 or 4-membered cycloalkoxy, 3 or            4-membered cycloalkyl, 3 or 4-membered heterocycloalkyl,            carboxy, aryl, cyano, halo, and heteroaryl, wherein aryl and            heteroaryl are optionally further substituted with one to            five substituents each independently selected from C₁-C₃            alkyl, C₁-C₃ haloalkyl, and halo; and    -   R⁵, R⁶, R⁷ and R⁸ are each independently selected from H, C₁-C₄        alkyl, C₁-C₄ haloalkyl, and halo.

Also provided is a pharmaceutical composition comprising a compound, ora pharmaceutically acceptable salt thereof, described herein and apharmaceutically acceptable carrier.

Also provided is a process for preparing a pharmaceutical compositioncomprising admixing a compound, or a pharmaceutically acceptable saltthereof, described herein and a pharmaceutically acceptable carrier.

Also provided is a method for treating a condition or disorder mediatedby at least one histone deacetylase in a patient in need thereofcomprising administering to the patient a therapeutically effectiveamount of a compound, or a pharmaceutically acceptable salt thereof,described herein.

DETAILED DESCRIPTION

As used in the present specification, the following words, phrases andsymbols are generally intended to have the meanings as set forth below,except to the extent that the context in which they are used indicatesotherwise.

A dash (“-”) that is not between two letters or symbols is used toindicate a point of attachment for a substituent. For example, —CONH₂ isattached through the carbon atom.

By “optional” or “optionally” is meant that the subsequently describedevent or circumstance may or may not occur, and that the descriptionincludes instances where the event or circumstance occurs and instancesin which it does not. For example, “optionally substituted alkyl”encompasses both “alkyl” and “substituted alkyl” as defined below. Itwill be understood by those skilled in the art, with respect to anygroup containing one or more substituents, that such groups are notintended to introduce any substitution or substitution patterns that aresterically impractical, synthetically non-feasible and/or inherentlyunstable.

“Alkyl” encompasses a straight chain and branched chain having theindicated number of carbon atoms, usually from 1 to 20 carbon atoms, forexample 1 to 8 carbon atoms, such as 1 to 6 carbon atoms. For exampleC₁-C₆ alkyl encompasses both straight and branched chain alkyl of from 1to 6 carbon atoms. Examples of alkyl groups include methyl, ethyl,propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, 2-pentyl,isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl, 3-methylpentyl, and thelike. Alkylene is another subset of alkyl, referring to the sameresidues as alkyl, but having two points of attachment. Alkylene groupswill usually have from 2 to 20 carbon atoms, for example 2 to 8 carbonatoms, such as from 2 to 6 carbon atoms. For example, C₀ alkyleneindicates a covalent bond and C₁ alkylene is a methylene group. When analkyl residue having a specific number of carbons is named, allgeometric isomers having that number of carbons are intended to beencompassed; thus, for example, “butyl” is meant to include n-butyl,sec-butyl, isobutyl and t-butyl; “propyl” includes n-propyl andisopropyl.

The term “alkylene” encompasses straight chain and branched chaindi-radical having the indicated number of carbon atoms, usually from 1to 20 carbon atoms, for example 1 to 8 carbon atoms, such as 1 to 6carbon atoms, or 1 to 4 carbon atoms. Examples of C₁-C₄ alkylene includemethylene, 1,1-ethyene, 1,2-ethylene, 1,1-propylene, 1,2-propylene,1,3-propylene, 1,1-butylene, 1,2-butylene, 1,3-butylene, 1,4-butylene,2-methyl-1,2-propylene, and 2-methyl-1,3-propylene.

By “alkoxy” is meant an alkyl group of the indicated number of carbonatoms attached through an oxygen bridge such as, for example, methoxy,ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, pentoxy,2-pentyloxy, isopentoxy, neopentoxy, hexoxy, 2-hexoxy, 3-hexoxy,3-methylpentoxy, and the like. Alkoxy groups will usually have from 1 to6 carbon atoms attached through the oxygen bridge.

“Aryl” indicates an aromatic carbon ring having the indicated number ofcarbon atoms, for example, 6 to 12 or 6 to 10 carbon atoms. Aryl groupsmay be monocyclic or polycyclic (e.g., bicyclic, tricyclic). In someinstances, both rings of a polycyclic aryl group are aromatic (e.g.,naphthyl). In other instances, polycyclic aryl groups may include anon-aromatic ring (e.g., cycloalkyl, cycloalkenyl, heterocycloalkyl,heterocycloalkenyl) fused to an aromatic ring, provided the polycyclicaryl group is bound to the parent structure via an atom in the aromaticring. Thus, a 1,2,3,4-tetrahydronaphthalen-5-yl group (wherein themoiety is bound to the parent structure via an aromatic carbon atom) isconsidered an aryl group, while 1,2,3,4-tetrahydronaphthalen-1-yl(wherein the moiety is bound to the parent structure via a non-aromaticcarbon atom) is not considered an aryl group. Similarly, a1,2,3,4-tetrahydroquinolin-8-yl group (wherein the moiety is bound tothe parent structure via an aromatic carbon atom) is considered an arylgroup, while 1,2,3,4-tetrahydroquinolin-1-yl group (wherein the moietyis bound to the parent structure via a non-aromatic nitrogen atom) isnot considered an aryl group. However, the term “aryl” does notencompass or overlap with “heteroaryl”, as defined herein, regardless ofthe point of attachment (e.g., both quinolin-5-yl and quinolin-2-yl areheteroaryl groups). In some instances, aryl is phenyl or naphthyl. Incertain instances, aryl is phenyl.

Bivalent radicals formed from substituted benzene derivatives and havingthe free valences at ring atoms are named as substituted phenyleneradicals. Bivalent radicals derived from univalent polycyclichydrocarbon radicals whose names end in “-yl” by removal of one hydrogenatom from the carbon atom with the free valence are named by adding“-idene” to the name of the corresponding univalent radical, e.g., anaphthyl group with two points of attachment is termed naphthylidene.

“Carboxy” indicates —C(O)OH.

“Cyano” refers to —CN.

“Cycloalkyl” indicates a non-aromatic, fully saturated carbocyclic ringhaving the indicated number of carbon atoms, for example, 3 to 10, or 3to 8, or 3 to 6 ring carbon atoms. Cycloalkyl groups may be monocyclicor polycyclic (e.g., bicyclic, tricyclic). Examples of cycloalkyl groupsinclude cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl andcyclohexyl, as well as bridged, spirocyclic, and caged ring groups(e.g., norbornane, bicyclo[2.2.2]octane). In addition, one ring of apolycyclic cycloalkyl group may be aromatic, provided the polycycliccycloalkyl group is bound to the parent structure via a non-aromaticcarbon. For example, a 1,2,3,4-tetrahydronaphthalen-1-yl group (whereinthe moiety is bound to the parent structure via a non-aromatic carbonatom) is a cycloalkyl group, while 1,2,3,4-tetrahydronaphthalen-5-yl(wherein the moiety is bound to the parent structure via an aromaticcarbon atom) is not considered a cycloalkyl group, i.e., it isconsidered an aryl group.

The term “cycloalkoxy” refers to “—O-cycloalkyl,” wherein cycloalkyl isas defined herein.

The term “halo” includes fluoro, chloro, bromo, and iodo, and the term“halogen” includes fluorine, chlorine, bromine, and iodine.

The term “haloalkyl” denotes an C₁₋₆ alkyl group wherein the alkyl issubstituted with one halogen up to fully substituted and a fullysubstituted C₁₋₆ haloalkyl can be represented by the formulaC_(n)L_(2n+1) wherein L is a halogen and “n” is 1, 2, 3 or 4; when morethan one halogen is present then they may be the same or different andselected from the group consisting of F, Cl, Br and I, such as F.Examples of C₁₋₄ haloalkyl groups include, but not limited to,fluoromethyl, difluoromethyl, trifluoromethyl, chlorodifluoromethyl,2,2,2-trifluoroethyl, pentafluoroethyl and the like.

The term “haloalkoxy” denotes a haloalkyl which is directly attached toan oxygen atom. Examples include, but not limited to, difluoromethoxy,trifluoromethoxy, 2,2,2-trifluoroethoxy, pentafluoroethoxy and the like.

“Heteroaryl” indicates an aromatic ring containing the indicated numberof atoms (e.g., 5 to 12, or 5 to 10 membered heteroaryl) made up of oneor more heteroatoms (e.g., 1, 2, 3 or 4 heteroatoms) selected from N, Oand S and with the remaining ring atoms being carbon.

Heteroaryl groups do not contain adjacent S and O atoms. In someembodiments, the total number of S and O atoms in the heteroaryl groupis not more than 2. In some embodiments, the total number of S and Oatoms in the heteroaryl group is not more than 1. Unless otherwiseindicated, heteroaryl groups may be bound to the parent structure by acarbon or nitrogen atom, as valency permits. For example, “pyridyl”includes 2-pyridyl, 3-pyridyl and 4-pyridyl groups, and “pyrrolyl”includes 1-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl groups. When nitrogen ispresent in a heteroaryl ring, it may, where the nature of the adjacentatoms and groups permits, exist in an oxidized state (i.e., N⁺—O⁻).Additionally, when sulfur is present in a heteroaryl ring, it may, wherethe nature of the adjacent atoms and groups permits, exist in anoxidized state (i.e., S⁺—O⁻ or SO₂). Heteroaryl groups may be monocyclicor polycyclic (e.g., bicyclic, tricyclic).

In some instances, a heteroaryl group is monocyclic. Examples includepyrrole, pyrazole, imidazole, triazole (e.g., 1,2,3-triazole,1,2,4-triazole), tetrazole, furan, isoxazole, oxazole, oxadiazole (e.g.,1,2,3-oxadiazole, 1,2,4-oxadiazole, 1,3,4-oxadiazole), thiophene,isothiazole, thiazole, thiadiazole (e.g., 1,2,3-thiadiazole,1,2,4-thiadiazole, 1,3,4-thiadiazole), pyridine, pyridazine, pyrimidine,pyrazine, triazine (e.g., 1,2,4-triazine, 1,3,5-triazine) and tetrazine.

In some instances, both rings of a polycyclic heteroaryl group arearomatic. Examples include indole, isoindole, indazole, benzoimidazole,benzotriazole, benzofuran, benzoxazole, benzoisoxazole, benzoxadiazole,benzothiophene, benzothiazole, benzoisothiazole, benzothiadiazole,1H-pyrrolo[2,3-b]pyridine, 1H-pyrazolo[3,4-b]pyridine,3H-imidazo[4,5-b]pyridine, 3H-[1,2,3]triazolo[4,5-b]pyridine,1H-pyrrolo[3,2-b]pyridine, 1H-pyrazolo[4,3-b]pyridine,1H-imidazo[4,5-b]pyridine, 1H-[1,2,3]triazolo[4,5-b]pyridine,1H-pyrrolo[2,3-c]pyridine, 1H-pyrazolo[3,4-c]pyridine,3H-imidazo[4,5-c]pyridine, 3H-[1,2,3]triazolo[4,5-c]pyridine,1H-pyrrolo[3,2-c]pyridine, 1H-pyrazolo[4,3-c]pyridine,1H-imidazo[4,5-c]pyridine, 1H-[1,2,3]triazolo[4,5-c]pyridine,furo[2,3-b]pyridine, oxazolo[5,4-b]pyridine, isoxazolo[5,4-b]pyridine,[1,2,3]oxadiazolo[5,4-b]pyridine, furo[3,2-b]pyridine,oxazolo[4,5-b]pyridine, isoxazolo[4,5-b]pyridine,[1,2,3]oxadiazolo[4,5-b]pyridine, furo[2,3-c]pyridine,oxazolo[5,4-c]pyridine, isoxazolo[5,4-c]pyridine,[1,2,3]oxadiazolo[5,4-c]pyridine, furo[3,2-c]pyridine,oxazolo[4,5-c]pyridine, isoxazolo[4,5-c]pyridine,[1,2,3]oxadiazolo[4,5-c]pyridine, thieno[2,3-b]pyridine,thiazolo[5,4-b]pyridine, isothiazolo[5,4-b]pyridine,[1,2,3]thiadiazolo[5,4-b]pyridine, thieno[3,2-b]pyridine,thiazolo[4,5-b]pyridine, isothiazolo[4,5-b]pyridine,[1,2,3]thiadiazolo[4,5-b]pyridine, thieno[2,3-c]pyridine,thiazolo[5,4-c]pyridine, isothiazolo[5,4-c]pyridine,[1,2,3]thiadiazolo[5,4-c]pyridine, thieno[3,2-c]pyridine,thiazolo[4,5-c]pyridine, isothiazolo[4,5-c]pyridine,[1,2,3]thiadiazolo[4,5-c]pyridine, quinoline, isoquinoline, cinnoline,quinazoline, quinoxaline, phthalazine, naphthyridine (e.g.,1,8-naphthyridine, 1,7-naphthyridine, 1,6-naphthyridine,1,5-naphthyridine, 2,7-naphthyridine, 2,6-naphthyridine),imidazo[1,2-a]pyridine, 1H-pyrazolo[3,4-d]thiazole,1H-pyrazolo[4,3-d]thiazole and imidazo[2,1-b]thiazole.

In other instances, polycyclic heteroaryl groups may include anon-aromatic ring (e.g., cycloalkyl, cycloalkenyl, heterocycloalkyl,heterocycloalkenyl) fused to a heteroaryl ring, provided the polycyclicheteroaryl group is bound to the parent structure via an atom in thearomatic ring. For example, a 4,5,6,7-tetrahydrobenzo[d]thiazol-2-ylgroup (wherein the moiety is bound to the parent structure via anaromatic carbon atom) is considered a heteroaryl group, while4,5,6,7-tetrahydrobenzo[d]thiazol-5-yl (wherein the moiety is bound tothe parent structure via a non-aromatic carbon atom) is not considered aheteroaryl group.

“Heterocycloalkyl” indicates a non-aromatic, fully saturated ring havingthe indicated number of atoms (e.g., 3 to 10, or 3 to 7, memberedheterocycloalkyl) made up of one or more heteroatoms (e.g., 1, 2, 3 or 4heteroatoms) selected from N, O and S and with the remaining ring atomsbeing carbon. Heterocycloalkyl groups may be monocyclic (i.e.,heteromonocyclic) or polycyclic (e.g., bicyclic (i.e., heterobicyclic),including spirocyclic and bridged ring systems). That is, the definitionof heterobicyclic encompasses a heteromonocyclic ring 1,1-disubstitutedwith a cycloalkyl or heteromonocyclic group, as well as a ring systemwherein a heteromonocyclic ring is 1,2- or 1,3-fused to anothercycloalkyl or heteromonocyclic ring (where a carbon or nitrogen atom canform the ring junction (where the structure is chemically feasible)), aswell as a ring system wherein a heteromonocyclic ring has a C₁-C₂ alkylbridge, as well as a ring system wherein a heteromonocyclic ring is1,2-fused to an aromatic or heteroaromatic ring, provided that themoiety is bound to the parent structure via a non-aromatic carbon ornitrogen atom.

Examples of monocyclic heterocycloalkyl (i.e., heteromonocyclic) groupsinclude oxiranyl, aziridinyl, azetidinyl, oxetanyl, pyrrolidinyl,imidazolidinyl, pyrazolidinyl, piperidinyl, piperazinyl, morpholinyl andthiomorpholinyl.

Examples of a C₆ heterobicyclyl group include3-azabicyclo[3.1.0]hexan-3-yl.

Examples of a C₈-C₁₀ heterobicyclyl group having an aromatic ringinclude indolin-1-yl, isoindolin-2-yl, 1,2,3,4-tetrahydroquinolin-2-yl,3,4-dihydroquinolin-1(2H)-yl, and 7,8-dihydro-1,6-naphthyridin-6(5H)-yl.

Examples of heterobicyclyl ring systems including a spirocycle include:1-oxa-5-azaspiro[3.3]heptan-5-yl, 1-oxa-6-azaspiro[3.3]heptan-6-yl,6-oxa-1-azaspiro[3.3]heptan-1-yl, 2-oxa-6-azaspiro[3.3]heptan-6-yl,1,5-diazaspiro[3.3]heptan-1-yl, 1,6-diazaspiro[3.3]heptan-6-yl,1,6-diazaspiro[3.3]heptan-1-yl, 2,6-diazaspiro[3.3]heptan-2-yl,1-oxa-5-azaspiro[3.4]octan-5-yl, 1-oxa-6-azaspiro[3.4]octan-6-yl,2-oxa-5-azaspiro[3.4]octan-5-yl, 2-oxa-6-azaspiro[3.4]octan-6-yl,1,5-diazaspiro[3.4]octan-5-yl, 1,6-diazaspiro[3.4]octan-6-yl,2,5-diazaspiro[3.4]octan-5-yl, 2,6-diazaspiro[3.4]octan-6-yl,1-oxa-5-azaspiro[3.5]nonan-5-yl, 1-oxa-6-azaspiro[3.5]nonan-6-yl,1-oxa-7-azaspiro[3.5]nonan-7-yl, 2-oxa-5-azaspiro[3.5]nonan-5-yl,2-oxa-6-azaspiro[3.5]nonan-6-yl, 2-oxa-7-azaspiro[3.5]nonan-7-yl,1,5-diazaspiro[3.5]nonan-5-yl, 1,6-diazaspiro[3.5]nonan-6-yl,1,7-diazaspiro[3.5]nonan-7-yl, 2,5-diazaspiro[3.5]nonan-5-yl,2,6-diazaspiro[3.5]nonan-6-yl, 2,7-diazaspiro[3.5]nonan-7-yl,1-oxa-5-azaspiro[3.6]decan-5-yl, 1-oxa-6-azaspiro[3.6]decan-6-yl,1-oxa-7-azaspiro[3.6]decan-7-yl, 2-oxa-5-azaspiro[3.6]decan-5-yl,2-oxa-6-azaspiro[3.6]decan-6-yl, 2-oxa-7-azaspiro[3.6]decan-7-yl,1,5-diazaspiro[3.6]decan-5-yl, 1,6-diazaspiro[3.6]decan-6-yl,1,7-diazaspiro[3.6]decan-7-yl 2,5-diazaspiro[3.6]decan-5-yl,2,6-diazaspiro[3.6]decan-6-yl, 2,7-diazaspiro[3.6]decan-7-yl.

Examples of heterobicyclyl ring systems including a C₁-C₄bridged-alkylene include 2-azabicyclo[2.2.1]heptan-2-yl,2-azabicyclo[3.2.1]octan-2-yl, 3-azabicyclo[3.2.1]octan-3-yl, and6-azabicyclo[3.2.1]octan-6-yl.

When nitrogen is present in a heterocycloalkyl ring, it may, where thenature of the adjacent atoms and groups permits, exist in an oxidizedstate (i.e., N⁺—O⁻). Examples include piperidinyl N-oxide andmorpholinyl-N-oxide. Additionally, when sulfur is present in aheterocycloalkyl ring, it may, where the nature of the adjacent atomsand groups permits, exist in an oxidized state (i.e., S⁺—O⁻ or —SO₂—).Examples include thiomorpholine S-oxide and thiomorpholine S,S-dioxide.

“Oxo” refers to (═O) or (O).

“Nitro” refers to —NO₂.

The term “substituted”, as used herein, means that any one or morehydrogens on the designated atom or group is replaced with a selectionfrom the indicated group, provided that the designated atom's normalvalence is not exceeded. When a substituent is oxo (i.e., ═O) then 2hydrogens on the atom are replaced. Combinations of substituents and/orvariables are permissible only if such combinations result in stablecompounds or useful synthetic intermediates. A stable compound or stablestructure is meant to imply a compound that is sufficiently robust tosurvive isolation from a reaction mixture, and subsequent formulation asan agent having at least practical utility. Unless otherwise specified,substituents are named into the core structure. For example, it is to beunderstood that when (cycloalkyl)alkyl is listed as a possiblesubstituent, the point of attachment of this substituent to the corestructure is in the alkyl portion.

The terms “substituted” alkyl (including without limitation C₁-C₄alkyl), cycloalkyl, aryl, heterocycloalkyl, and heteroaryl, unlessotherwise expressly defined, refer respectively to alkyl, cycloalkyl,aryl, heterocycloalkyl, and heteroaryl wherein one or more (such as upto 5, for example, up to 3) hydrogen atoms are replaced by a substituentindependently chosen from —R^(a), —OR^(b), —O(C₁-C₂ alkyl)O— (e.g.,methylenedioxy-), —SR^(b), guanidine (—NHC(═NH)NH₂), guanidine whereinone or more of the guanidine hydrogens are replaced with a C₁-C₄alkylgroup, —NR^(b)R^(c), halo, cyano, oxo (as a substituent forheterocycloalkyl), nitro, —COR^(b), —CO₂R^(b), —CONR^(b)R^(c),—OCOR^(b), —OCO₂R^(a), —OCONR^(b)R^(c), —NR^(c)COR^(b), —NR^(c)CO₂R^(a),—NR^(c)CONR^(b)R^(c), —SOR^(a), —SO₂R^(a), —SO₂NR^(b)R^(c), and—NR^(c)SO₂R^(a),

where R^(a) is chosen from C₁-C₆ alkyl, cycloalkyl, aryl,heterocycloalkyl, and heteroaryl;

R^(b) is chosen from H, C₁-C₆ alkyl, aryl, and heteroaryl; and

R^(c) is chosen from hydrogen and C₁-C₄ alkyl; or

R^(b) and R^(c), and the nitrogen to which they are attached, form aheterocycloalkyl group; and where each C₁-C₆ alkyl, cycloalkyl, aryl,heterocycloalkyl, and heteroaryl is optionally substituted with one ormore, such as one, two, or three, substituents independently selectedfrom C₁-C₄ alkyl, C₃-C₆ cycloalkyl, aryl, heteroaryl, aryl-C₁-C₄ alkyl-,heteroaryl-C₁-C₄ alkyl-, C₁-C₄ haloalkyl-, —OC₁-C₄ alkyl, —OC₁-C₄alkylphenyl, —C₁-C₄ alkyl-OH, —C₁-C₄ alkyl-O—C₁-C₄ alkyl, —OC₁-C₄haloalkyl, halo, —OH, —NH₂, —C₁-C₄ alkyl-NH₂, —N(C₁-C₄ alkyl)(C₁-C₄alkyl), —NH(C₁-C₄ alkyl), —N(C₁-C₄ alkyl)(C₁-C₄ alkylphenyl), —NH(C₁-C₄alkylphenyl), cyano, nitro, oxo (as a substituent for heteroaryl),—CO₂H, —C(O)OC₁-C₄ alkyl, —CON(C₁-C₄ alkyl)(C₁-C₄ alkyl), —CONH(C₁-C₄alkyl), —CONH₂, —NHC(O)(C₁-C₄ alkyl), —NHC(O)(phenyl), —N(C₁-C₄alkyl)C(O)(C₁-C₄ alkyl), —N(C₁-C₄ alkyl)C(O)(phenyl), —C(O)C₁-C₄ alkyl,—C(O)C₁-C₄ phenyl, —C(O)C₁-C₄ haloalkyl, —OC(O)C₁-C₄ alkyl, —SO₂(C₁-C₄alkyl), —SO₂(phenyl), —SO₂(C₁-C₄ haloalkyl), —SO₂NH₂, —SO₂NH(C₁-C₄alkyl), —SO₂NH(phenyl), —NHSO₂(C₁-C₄ alkyl), —NHSO₂(phenyl), and—NHSO₂(C₁-C₄ haloalkyl).

Compounds described herein include, but are not limited to, theiroptical isomers, racemates, and other mixtures thereof. In thosesituations, the single enantiomers or diastereomers, i.e., opticallyactive forms, can be obtained by asymmetric synthesis or by resolutionof the racemates. Resolution of the racemates can be accomplished, forexample, by conventional methods such as crystallization in the presenceof a resolving agent, or chromatography, using, for example a chiralhigh-pressure liquid chromatography (HPLC) or supercritical fluidchromatograph (SFC) column. In addition, such compounds include Z- andE-forms (or cis- and trans-forms) of compounds with carbon-carbon doublebonds. Where compounds described herein exist in various tautomericforms, the term “compound” is intended to include all tautomeric formsof the compound. Such compounds also include crystal forms includingpolymorphs and clathrates. Similarly, the term “salt” is intended toinclude all tautomeric forms and crystal forms of the compound.

Where a configuration of a single diastereomer is not known theconfiguration has been denoted, for example, as D1 (diastereomer 1) andD2 (diastereomer 2) and the unknown chiral center(s) labeled with anasterisk. For example, D1N—((R)-1-((abs)-3-(difluoromethoxy)piperidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamideand D2N—((R)-1-((abs)-3-(difluoromethoxy)piperidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamideare single diastereomers for which the configuration at one chiralcenter is known absolutely (with configuration drawn accordingly) andthe configuration at the second chiral center is absolute but unknown(drawn as a bond with an asterisk), i.e. opposite configuration at theunknown center for D1 versus D2.

Where a single isomer has been isolated for a compound with three chiralcenters where one stereocenter is known, and the absolute configurationof the other two centers are unknown but the relative configurationknown to be cis, e.g. a homochiral azabicycloheptanyl ring system, thecompound has been drawn where the unknown chiral center(s) are labeledwith an asterisk, and named accordingly, i.e. D1:N—((R)-1-((abs-1,5-cis)-6-azabicyclo[3.2.0]heptan-6-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;and D2:N—((R)-1-((abs-1,5-cis)-6-azabicyclo[3.2.0]heptan-6-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide.

“Pharmaceutically acceptable salts” include, but are not limited tosalts with inorganic acids, such as hydrochloride, phosphate,diphosphate, hydrobromide, sulfate, sulfinate, nitrate, and like salts;as well as salts with an organic acid, such as malate, maleate,fumarate, tartrate, succinate, citrate, acetate, lactate,methanesulfonate, p-toluenesulfonate, 2-hydroxyethylsulfonate, benzoate,salicylate, stearate, and alkanoate such as acetate, HOOC—(CH₂)_(q)—COOHwhere q is 0-4, and like salts. Similarly, pharmaceutically acceptablecations include, but are not limited to sodium, potassium, calcium,aluminum, lithium, and ammonium.

In addition, if the compounds described herein are obtained as an acidaddition salt, the free base can be obtained by basifying a solution ofthe acid salt. Conversely, if the product is a free base, an additionsalt, particularly a pharmaceutically acceptable addition salt, may beproduced by dissolving the free base in a suitable organic solvent andtreating the solution with an acid, in accordance with conventionalprocedures for preparing acid addition salts from base compounds. Thoseskilled in the art will recognize various synthetic methodologies thatmay be used to prepare non-toxic free base or non-toxic pharmaceuticallyacceptable addition salts.

As used herein the terms “group”, “radical” or “fragment” are synonymousand are intended to indicate functional groups or fragments of moleculesattachable to a bond or other fragments of molecules.

The term “active agent” is used to indicate a compound or apharmaceutically acceptable salt thereof which has biological activity.In some embodiments, an “active agent” is a compound or pharmaceuticallyacceptable salt thereof having pharmaceutical utility. For example anactive agent may be an anti-neurodegenerative therapeutic.

The term “therapeutically effective amount” means an amount effective,when administered to a human or non-human patient, to provide atherapeutic benefit such as amelioration of symptoms, slowing of diseaseprogression, or prevention of disease e.g., a therapeutically effectiveamount may be an amount sufficient to decrease the symptoms of a diseaseresponsive to inhibition of HDAC activity.

As used herein, the terms “histone deacetylase” and “HDAC” are intendedto refer to any one of a family of enzymes that remove N^(ε)-acetylgroups from the ε-amino groups of lysine residues of a protein (forexample, a histone, or tubulin). Unless otherwise indicated by context,the term “histone” is meant to refer to any histone protein, includingH1, H2A, H2B, H3, H4, and H5, from any species. In some embodiments, thehistone deacetylase is a human HDAC, including, but not limited to,HDAC-4, HDAC-5, HDAC-6, HDAC-7, HDAC-9, and HDAC-10. In someembodiments, at least one histone deacetylase is selected from HDAC-4,HDAC-5, HDAC-7, and HDAC-9. In some embodiments, the histone deacetylaseis a class IIa HDAC. In some embodiments, the histone deacetylase isHDAC-4. In some embodiments, the histone deacetylase is HDAC-5. In someembodiments, the histone deacetylase is derived from a protozoal orfungal source.

The terms “histone deacetylase inhibitor” and “inhibitor of histonedeacetylase” are intended to mean a compound, or a pharmaceuticallyacceptable salt thereof, described herein which is capable ofinteracting with a histone deacetylase and inhibiting its enzymaticactivity.

The term “a condition or disorder mediated by HDAC” or “a condition ordisorder mediated by histone deacetylase” as used herein refers to acondition or disorder in which HDAC and/or the action of HDAC isimportant or necessary, e.g., for the onset, progress, expression, etc.of that condition, or a condition which is known to be treated by HDACinhibitors (such as, trichostatin A).

The term “effect” describes a change or an absence of a change in cellphenotype or cell proliferation. “Effect” can also describe a change oran absence of a change in the catalytic activity of HDAC. “Effect” canalso describe a change or an absence of a change in an interactionbetween HDAC and a natural binding partner.

The term “inhibiting histone deacetylase enzymatic activity” or“inhibiting histone deacetylase” is intended to mean reducing theability of a histone deacetylase to remove an acetyl group from aprotein, such as but not limited to a histone or tubulin. Theconcentration of inhibitor which reduces the activity of a histonedeacetylase to 50% of that of the uninhibited enzyme is determined asthe IC₅₀ value. In some embodiments, such reduction of histonedeacetylase activity is at least 50%, such as at least about 75%, forexample, at least about 90%. In some embodiments, histone deacetylaseactivity is reduced by at least 95%, such as by at least 99%. In someembodiments, the compounds and pharmaceutical acceptable salts thereofdescribed herein have an IC₅₀ value less than 100 nanomolar. In someembodiments, the compounds and pharmaceutical acceptable salts thereofdescribed herein have an IC₅₀ value from 100 nanomolar to 1 micromolar.In some embodiments, the compounds and pharmaceutical acceptable saltsthereof described herein have an IC₅₀ value from 1 to 25 micromolar.

In some embodiments, such inhibition is specific, i.e., the histonedeacetylase inhibitor reduces the ability of a histone deacetylase toremove an acetyl group from a protein at a concentration that is lowerthan the concentration of the inhibitor that is required to produceanother, unrelated biological effect. In some embodiments, theconcentration of the inhibitor required for histone deacetylaseinhibitory activity is at least 2-fold lower, such as at least 5-foldlower, for example, at least 10-fold lower, such as at least 20-foldlower than the concentration required to produce an unrelated biologicaleffect.

“Treatment” or “treating” means any treatment of a disease state in apatient, including

a) preventing the disease, that is, causing the clinical symptoms of thedisease not to develop;

b) inhibiting the disease;

c) slowing or arresting the development of clinical symptoms; and/or

d) relieving the disease, that is, causing the regression of clinicalsymptoms.

“Subject” or “patient’ refers to an animal, such as a mammal, that hasbeen or will be the object of treatment, observation or experiment. Themethods described herein may be useful in both human therapy andveterinary applications. In some embodiments, the subject is a mammal;and in some embodiments the subject is human.

It is appreciated that certain features described herein, which are, forclarity, described in the context of separate embodiments, may also beprovided in combination in a single embodiment. Conversely, variousfeatures described herein, which are, for brevity, described in thecontext of a single embodiment, may also be provided separately or inany suitable subcombination. All combinations of the embodimentspertaining to the chemical groups represented by the variables containedwithin Formula I, are specifically embraced by herein just as if eachand every combination was individually and explicitly recited, to theextent that such combinations embrace compounds that result in stablecompounds (i.e., compounds that can be isolated, characterized andtested for biological activity). In addition, all subcombinations of thechemical groups listed in the embodiments describing such variables, aswell as all subcombinations of uses and medical indications describedherein, such as those conditions or disorders mediated by HDAC, are alsospecifically embraced herein just as if each and every subcombination ofchemical groups and subcombination of uses and medical indications wasindividually and explicitly recited herein. In addition, someembodiments include every combination of one or more additional agentsdisclosed herein just as if each and every combination was individuallyand explicitly recited.

Provided is a compound of Formula I:

or a pharmaceutically acceptable salt thereof;

wherein:

-   -   W is N or CR⁵; X is N or CR⁶; Y is N or CR⁷; and Z is N or CR⁸;        provided not more than two of W, X, Y, and Z are N;    -   R¹ is selected from H and C₁-C₃ alkyl;    -   R² is C₂-C₃ alkylene optionally substituted with C₁-C₂ haloalkyl        or 3 or 4-membered cycloalkyl;    -   R³ and R⁴, together with the nitrogen atom to which they are        attached, form:        -   a 4, 5, 6, or 7-membered heteromonocyclic group, or        -   a 6, 7, 8, 9, or 10-membered heterobicyclic group,        -   each of which is optionally substituted with one to five            substituents each independently selected from: C₁-C₃ alkoxy,            C₁-C₃ alkyl, C₁-C₃ haloalkoxy, C₁-C₃ haloalkyl, 3 or            4-membered cycloalkoxy, 3 or 4-membered cycloalkyl, 3 or            4-membered heterocycloalkyl, carboxy, aryl, cyano, halo, and            heteroaryl, wherein aryl and heteroaryl are optionally            further substituted with one to five substituents each            independently selected from C₁-C₃ alkyl, C₁-C₃ haloalkyl,            and halo,    -   provided that if R³ and R⁴, together with the nitrogen atom to        which they are attached,    -   form a 5 or 6-membered heteromonocyclic group, then:        -   i) R² is C₂-C₃ alkylene substituted with C₁-C₂ haloalkyl or            3 or 4-membered cycloalkyl, or        -   ii) R⁸ is halo or C₁-C₃ alkyl, or        -   iii) the 5 or 6-membered heteromonocyclic group is            substituted with one to five substituents each independently            selected from: C₁-C₃ alkoxy, C₁-C₃ alkyl, C₁-C₃ haloalkoxy,            C₁-C₃ haloalkyl, 3 or 4-membered cycloalkoxy, 3 or            4-membered cycloalkyl, 3 or 4-membered heterocycloalkyl,            carboxy, aryl, cyano, halo, and heteroaryl, wherein aryl and            heteroaryl are optionally further substituted with one to            five substituents each independently selected from C₁-C₃            alkyl, C₁-C₃ haloalkyl, and halo; and    -   R⁵, R⁶, R⁷ and R⁸ are each independently selected from H, C₁-C₄        alkyl, C₁-C₄ haloalkyl, and halo.

In some embodiments, R¹ is selected from H and methyl. In someembodiments, R¹ is H.

In some embodiments, R² is C₂-C₃ alkylene optionally substituted with 3or 4-membered cycloalkyl. In some embodiments, R² is chosen from—(CH₂)₂— and —CH(CH₃)CH₂—.

In some embodiments, R⁵ is H.

In some embodiments, R⁶ is H.

In some embodiments, R⁷ is H.

In some embodiments, R⁸ is selected from H, halo, and methyl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 4 or 7-membered heteromonocyclic groupoptionally substituted with one to five substituents each independentlyselected from: C₁-C₃ alkoxy, C₁-C₃ alkyl, C₁-C₃ haloalkoxy, C₁-C₃haloalkyl, 3 or 4-membered cycloalkoxy, 3 or 4-membered cycloalkyl, 3 or4-membered heterocycloalkyl, carboxy, aryl, cyano, halo, and heteroaryl,wherein aryl and heteroaryl are optionally further substituted with oneto five substituents each independently selected from C₁-C₃ alkyl, C₁-C₃haloalkyl, and halo.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 4 or 7-membered heteromonocyclic groupoptionally substituted with one to five substituents each independentlyselected from: C₁-C₃ alkoxy, C₁-C₃ alkyl, C₁-C₃ haloalkoxy, C₁-C₃haloalkyl, 3 or 4-membered cycloalkoxy, 3 or 4-membered cycloalkyl, 3 or4-membered heterocycloalkyl, aryl, and heteroaryl, wherein aryl andheteroaryl are optionally further substituted with one to fivesubstituents each independently selected from C₁-C₃ alkyl, C₁-C₃haloalkyl, and halo.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 4 or 7-membered heteromonocyclic groupoptionally substituted with one to five substituents each independentlyselected from: C₁-C₃ alkyl, C₁-C₃ haloalkoxy, and aryl, wherein aryl isoptionally further substituted with one to five substituents eachindependently selected from C₁-C₃ alkyl, C₁-C₃ haloalkyl, and halo.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 4 or 7-membered heteromonocyclic groupoptionally substituted with one to five substituents each independentlyselected from: C₁-C₃ alkyl, C₁-C₃ haloalkoxy, and phenyl, wherein phenylis optionally further substituted with one to five substituents eachindependently selected from C₁-C₃ alkyl, C₁-C₃ haloalkyl, and halo.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 4 or 7-membered heteromonocyclic groupoptionally substituted with one to five substituents each independentlyselected from: methyl, difluoromethoxy, and phenyl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 4 or 7-membered heteromonocyclic groupselected from 3-phenylazetidin-1-yl, and azepan-1-yl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic group and R²is C₂-C₃ alkylene substituted with C₁-C₂ haloalkyl or 3 or 4-memberedcycloalkyl, such as R² is C₂-C₃ alkylene substituted with cyclopropyl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic groupoptionally substituted with one to five substituents each independentlyselected from: C₁-C₃ alkoxy, C₁-C₃ alkyl, C₁-C₃ haloalkoxy, C₁-C₃haloalkyl, 3 or 4-membered cycloalkoxy, 3 or 4-membered cycloalkyl, 3 or4-membered heterocycloalkyl, carboxy, aryl, cyano, halo, and heteroaryl,wherein aryl and heteroaryl are optionally further substituted with oneto five substituents each independently selected from C₁-C₃ alkyl, C₁-C₃haloalkyl, and halo and R² is C₂-C₃ alkylene substituted with C₁-C₂haloalkyl or 3 or 4-membered cycloalkyl, such as R² is C₂-C₃ alkylenesubstituted with cyclopropyl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic groupoptionally substituted with one to five substituents each independentlyselected from: C₁-C₃ alkoxy, C₁-C₃ alkyl, C₁-C₃ haloalkoxy, C₁-C₃haloalkyl, 3 or 4-membered cycloalkoxy, 3 or 4-membered cycloalkyl, 3 or4-membered heterocycloalkyl, aryl, and heteroaryl, wherein aryl andheteroaryl are optionally further substituted with one to fivesubstituents each independently selected from C₁-C₃ alkyl, C₁-C₃haloalkyl, and halo and R² is C₂-C₃ alkylene substituted with C₁-C₂haloalkyl or 3 or 4-membered cycloalkyl, such as R² is C₂-C₃ alkylenesubstituted with cyclopropyl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic groupoptionally substituted with one to five substituents each independentlyselected from: C₁-C₃ alkyl, C₁-C₃ haloalkoxy, and aryl, wherein aryl isoptionally further substituted with one to five substituents eachindependently selected from C₁-C₃ alkyl, C₁-C₃ haloalkyl, and halo andR² is C₂-C₃ alkylene substituted with C₁-C₂ haloalkyl or 3 or 4-memberedcycloalkyl, such as R² is C₂-C₃ alkylene substituted with cyclopropyl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic groupoptionally substituted with one to five substituents each independentlyselected from: C₁-C₃ alkyl, C₁-C₃ haloalkoxy, and phenyl, wherein phenylis optionally further substituted with one to five substituents eachindependently selected from C₁-C₃ alkyl, C₁-C₃ haloalkyl, and halo andR² is C₂-C₃ alkylene substituted with C₁-C₂ haloalkyl or 3 or 4-memberedcycloalkyl, such as R² is C₂-C₃ alkylene substituted with cyclopropyl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic groupoptionally substituted with one to five substituents each independentlyselected from: methyl, difluoromethoxy, and phenyl and R² is C₂-C₃alkylene substituted with C₁-C₂ haloalkyl or 3 or 4-membered cycloalkyl,such as R² is C₂-C₃ alkylene substituted with cyclopropyl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic groupselected from pyrrolidin-1-yl, and piperidin-1-yl, each of which isoptionally substituted with one substituent selected from: methyl,difluoromethoxy, and phenyl and R² is C₂-C₃ alkylene substituted withC₁-C₂ haloalkyl or 3 or 4-membered cycloalkyl, such as R² is C₂-C₃alkylene substituted with cyclopropyl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic groupselected from pyrrolidin-1-yl, 2-methyl-pyrrolidin-1-yl, and3-(difluoromethoxy)piperidin-1-yl and R² is C₂-C₃ alkylene substitutedwith C₁-C₂ haloalkyl or 3 or 4-membered cycloalkyl, such as R² is C₂-C₃alkylene substituted with cyclopropyl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic group and R⁸is halo or C₁-C₃ alkyl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic groupoptionally substituted with one to five substituents each independentlyselected from: C₁-C₃ alkoxy, C₁-C₃ alkyl, C₁-C₃ haloalkoxy, C₁-C₃haloalkyl, 3 or 4-membered cycloalkoxy, 3 or 4-membered cycloalkyl, 3 or4-membered heterocycloalkyl, carboxy, aryl, cyano, halo, and heteroaryl,wherein aryl and heteroaryl are optionally further substituted with oneto five substituents each independently selected from C₁-C₃ alkyl, C₁-C₃haloalkyl, and halo and R⁸ is halo or C₁-C₃ alkyl, e.g., methyl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic groupoptionally substituted with one to five substituents each independentlyselected from: C₁-C₃ alkoxy, C₁-C₃ alkyl, C₁-C₃ haloalkoxy, C₁-C₃haloalkyl, 3 or 4-membered cycloalkoxy, 3 or 4-membered cycloalkyl, 3 or4-membered heterocycloalkyl, aryl, and heteroaryl, wherein aryl andheteroaryl are optionally further substituted with one to fivesubstituents each independently selected from C₁-C₃ alkyl, C₁-C₃haloalkyl, and halo and R⁸ is halo or C₁-C₃ alkyl, e.g., methyl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic groupoptionally substituted with one to five substituents each independentlyselected from: C₁-C₃ alkyl, C₁-C₃ haloalkoxy, and aryl, wherein aryl isoptionally further substituted with one to five substituents eachindependently selected from C₁-C₃ alkyl, C₁-C₃ haloalkyl, and halo andR⁸ is halo or C₁-C₃ alkyl, e.g., methyl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic groupoptionally substituted with one to five substituents each independentlyselected from: C₁-C₃ alkyl, C₁-C₃ haloalkoxy, and phenyl, wherein phenylis optionally further substituted with one to five substituents eachindependently selected from C₁-C₃ alkyl, C₁-C₃ haloalkyl, and halo andR⁸ is halo or C₁-C₃ alkyl, e.g., methyl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic groupoptionally substituted with one to five substituents each independentlyselected from: methyl, difluoromethoxy, and phenyl and R⁸ is halo orC₁-C₃ alkyl, e.g., methyl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic groupselected from pyrrolidin-1-yl, and piperidin-1-yl, each of which isoptionally substituted with one substituent selected from: methyl,difluoromethoxy, and phenyl and R⁸ is halo or C₁-C₃ alkyl, e.g., methyl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic groupselected from pyrrolidin-1-yl, 2-methyl-pyrrolidin-1-yl, and3-(difluoromethoxy)piperidin-1-yl and R⁸ is halo or C₁-C₃ alkyl, e.g.,methyl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic groupsubstituted with one to five substituents each independently selectedfrom: C₁-C₃ alkoxy, C₁-C₃ alkyl, C₁-C₃ haloalkoxy, C₁-C₃ haloalkyl, 3 or4-membered cycloalkoxy, 3 or 4-membered cycloalkyl, 3 or 4-memberedheterocycloalkyl, carboxy, aryl, cyano, halo, and heteroaryl, whereinaryl and heteroaryl are optionally further substituted with one to fivesubstituents each independently selected from C₁-C₃ alkyl, C₁-C₃haloalkyl, and halo.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic groupsubstituted with one to five substituents each independently selectedfrom: C₁-C₃ alkyl, C₁-C₃ haloalkoxy, and aryl, wherein aryl isoptionally further substituted with one to five substituents eachindependently selected from C₁-C₃ alkyl, C₁-C₃ haloalkyl, and halo.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic groupsubstituted with one to five substituents each independently selectedfrom: C₁-C₃ alkyl, C₁-C₃ haloalkoxy, and phenyl, wherein phenyl isoptionally further substituted with one to five substituents eachindependently selected from C₁-C₃ alkyl, C₁-C₃ haloalkyl, and halo.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic groupsubstituted with one to five substituents each independently selectedfrom: methyl, difluoromethoxy, and phenyl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic groupselected from pyrrolidin-1-yl, and piperidin-1-yl, each of which issubstituted with one substituent selected from: methyl, difluoromethoxy,and phenyl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic groupselected from 2-methyl-pyrrolidin-1-yl, and3-(difluoromethoxy)piperidin-1-yl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a (2S)-methyl-pyrrolidin-1-yl.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 6, 7, 8, 9, or 10-membered heterobicyclicgroup, which is optionally substituted with one to five substituentseach independently selected from: C₁-C₃ alkoxy, C₁-C₃ alkyl, C₁-C₃haloalkoxy, C₁-C₃ haloalkyl, 3 or 4-membered cycloalkoxy, 3 or4-membered cycloalkyl, 3 or 4-membered heterocycloalkyl, carboxy, aryl,cyano, halo, and heteroaryl, wherein aryl and heteroaryl are optionallyfurther substituted with one to five substituents each independentlyselected from C₁-C₃ alkyl, C₁-C₃ haloalkyl, and halo.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 6, 7, 8, 9, or 10-membered heterobicyclicgroup selected from:

-   1-azaspiro[3.3]heptan-1-yl,-   3-azabicyclo[3.1.0]hexan-3-yl,-   3-azabicyclo[3.2.0]heptan-3-yl,-   3-azabicyclo[3.2.1]heptan-3-yl,-   3-azabicyclo[3.2.1]octan-3-yl,-   3,4-dihydroisoquinolin-2(1H)-yl,-   3,4-dihydro-2,7-naphthyridin-2(1H)-yl,-   5-azaspiro[2.4]heptan-5-yl,-   5-azaspiro[2.5]octan-5-yl,-   6-azaspiro[2.5]octan-6-yl,-   1-azaspiro[3.3]heptan-1-yl,-   6-azabicyclo[3.2.0]heptan-6-yl,-   isoindolin-2-yl, and-   octahydrocyclopenta[c]pyrrol-2-yl, and    -   each of which is optionally substituted with one to five        substituents each independently selected from: C₁-C₃ alkoxy,        C₁-C₃ alkyl, C₁-C₃ haloalkoxy, C₁-C₃ haloalkyl, 3 or 4-membered        cycloalkoxy, 3 or 4-membered cycloalkyl, 3 or 4-membered        heterocycloalkyl, carboxy, aryl, cyano, halo, and heteroaryl,        wherein aryl and heteroaryl are optionally further substituted        with one to five substituents each independently selected from        C₁-C₃ alkyl, C₁-C₃ haloalkyl, and halo.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 6, 7, 8, 9, or 10-membered heterobicyclicgroup selected from:

-   1-azaspiro[3.3]heptan-1-yl,-   3-azabicyclo[3.1.0]hexan-3-yl,-   3-azabicyclo[3.2.0]heptan-3-yl,-   3-azabicyclo[3.2.1]heptan-3-yl,-   3-azabicyclo[3.2.1]octan-3-yl,-   3,4-dihydroisoquinolin-2(1H)-yl,-   3,4-dihydro-2,7-naphthyridin-2(1H)-yl,-   5-azaspiro[2.4]heptan-5-yl,-   5-azaspiro[2.5]octan-5-yl,-   6-azaspiro[2.5]octan-6-yl,-   1-azaspiro[3.3]heptan-1-yl,-   6-azabicyclo[3.2.0]heptan-6-yl,-   isoindolin-2-yl, and-   octahydrocyclopenta[c]pyrrol-2-yl, and    -   each of which is optionally substituted with one or two        substituents each independently selected from: C₁-C₃ alkoxy,        C₁-C₃ alkyl, C₁-C₃ haloalkoxy, C₁-C₃ haloalkyl, and carboxy.

In some embodiments, R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 6, 7, 8, 9, or 10-membered heterobicyclicgroup selected from:

-   1-azaspiro[3.3]heptan-1-yl,-   3-azabicyclo[3.1.0]hexan-3-yl,-   3-azabicyclo[3.2.0]heptan-3-yl,-   3-azabicyclo[3.2.1]heptan-3-yl,-   3-azabicyclo[3.2.1]octan-3-yl,-   3,4-dihydroisoquinolin-2(1H)-yl,-   3,4-dihydro-2,7-naphthyridin-2(1H)-yl,-   5-azaspiro[2.4]heptan-5-yl,-   5-azaspiro[2.5]octan-5-yl,-   6-azaspiro[2.5]octan-6-yl,-   1-azaspiro[3.3]heptan-1-yl,-   6-azabicyclo[3.2.0]heptan-6-yl,-   isoindolin-2-yl, and-   octahydrocyclopenta[c]pyrrol-2-yl, and    -   each of which is optionally substituted with one or two        substituents each independently selected from C₁-C₃ alkyl.

Also provided is a compound of Formula II, or a pharmaceuticallyacceptable salt thereof,

wherein R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are as described herein.

Also provided is a compound of Formula III, or a pharmaceuticallyacceptable salt thereof,

wherein R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are as described herein.

Also provided is a compound of Formula IV, or a pharmaceuticallyacceptable salt thereof, of Formula VI, or a pharmaceutically acceptablesalt thereof,

wherein R³ and R⁴ are as described herein.

Also provided is a compound of Formula V, or a pharmaceuticallyacceptable salt thereof,

wherein R¹, R², R³, R⁴, R⁵, R⁷, and R⁸ are as described herein.

Also provided is a compound of Formula VI, or a pharmaceuticallyacceptable salt thereof,

wherein R¹, R³, R⁴, R⁵, R⁷, and R⁸ are as described herein.

Also provided is a compound of Formula VII, or a pharmaceuticallyacceptable salt thereof,

wherein R³ and R⁴ are as described herein.

Also provided is a compound selected from:

-   (2S)-2-Methyl-1-((R)-2-(3-methyl-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamido)propyl)pyrrolidin-1-ium    formate;-   N-(2-(3-Azabicyclo[3.2.1]octan-3-yl)ethyl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (R)—N-(1-(Isoindolin-2-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   N-((2R)-1-(3-Azabicyclo[3.1.0]hexan-3-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (S)—N-(1-Cyclopropyl-2-(pyrrolidin-1-yl)ethyl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (R)—N-(1-Cyclopropyl-2-(pyrrolidin-1-yl)ethyl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (R)—N-(1-(3,4-Dihydroisoquinolin-2(1H)-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (R)—N-(1-(3-Phenylazetidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   D1:    N—((R)-1-((abs)-3-(Difluoromethoxy)piperidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   D2:    N—((R)-1-((abs)-3-(Difluoromethoxy)piperidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (R)—N-(1-(5-Azaspiro[2.5]octan-5-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (R)—N-(1-(6-Azaspiro[2.5]octan-6-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (R)—N-(1-(Azepan-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (R)—N-(1-(1-Azaspiro[3.3]heptan-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (R)—N-(1-(5-Azaspiro[2.4]heptan-5-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   N—((R)-1-(3-Azabicyclo[3.2.1]octan-3-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   2-((R)-2-(4-(5-(Trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamido)propyl)octahydrocyclopenta[c]pyrrol-2-ium    formate;-   (R)—N-(1-(3,4-dihydro-2,7-naphthyridin-2(1H)-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   N-((2R)-1-(3-Azabicyclo[3.2.0]heptan-3-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   D1:    N—((R)-1-((abs-1,5-cis)-6-Azabicyclo[3.2.0]heptan-6-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   D2:    N—((R)-1-((abs-1,5-cis)-6-Azabicyclo[3.2.0]heptan-6-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   N-((2R)-1-(6,6-Dimethyl-3-azabicyclo[3.1.0]hexan-3-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (2S)-1-((R)-2-(3-Fluoro-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamido)propyl)-2-methylpyrrolidin-1-ium    formate; and-   N—((R)-1-((S)-2-Methylpyrrolidin-1-yl)propan-2-yl)-5-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)picolinamide,    or a pharmaceutically acceptable salt thereof.

Also provided is a compound selected from:

-   (2S)-2-Methyl-1-((R)-2-(3-methyl-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamido)propyl)pyrrolidin-1-ium    formate;-   N-(2-(3-Azabicyclo[3.2.1]octan-3-yl)ethyl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (R)—N-(1-(Isoindolin-2-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   N-((2R)-1-(3-Azabicyclo[3.1.0]hexan-3-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (S)—N-(1-Cyclopropyl-2-(pyrrolidin-1-yl)ethyl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (R)—N-(1-Cyclopropyl-2-(pyrrolidin-1-yl)ethyl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (R)—N-(1-(3,4-Dihydroisoquinolin-2(1H)-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (R)—N-(1-(3-Phenylazetidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   D1:    N—((R)-1-((abs)-3-(Difluoromethoxy)piperidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   D2:    N—((R)-1-((abs)-3-(Difluoromethoxy)piperidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (R)—N-(1-(5-Azaspiro[2.5]octan-5-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (R)—N-(1-(6-Azaspiro[2.5]octan-6-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (R)—N-(1-(Azepan-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (R)—N-(1-(1-Azaspiro[3.3]heptan-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (R)—N-(1-(5-Azaspiro[2.4]heptan-5-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   N—((R)-1-(3-Azabicyclo[3.2.1]octan-3-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   2-((R)-2-(4-(5-(Trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamido)propyl)octahydrocyclopenta[c]pyrrol-2-ium    formate;-   (R)—N-(1-(3,4-dihydro-2,7-naphthyridin-2(1H)-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   N-((2R)-1-(3-Azabicyclo[3.2.0]heptan-3-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   D1:    N—((R)-1-((abs-1,5-cis)-6-Azabicyclo[3.2.0]heptan-6-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   D2:    N—((R)-1-((abs-1,5-cis)-6-Azabicyclo[3.2.0]heptan-6-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   N-((2R)-1-(6,6-Dimethyl-3-azabicyclo[3.1.0]hexan-3-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   (2S)-1-((R)-2-(3-Fluoro-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamido)propyl)-2-methylpyrrolidin-1-ium    formate;-   N—((R)-1-((S)-2-Methylpyrrolidin-1-yl)propan-2-yl)-5-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)picolinamide;-   3-methyl-N—((R)-1-((S)-2-methylpyrrolidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   N-((2R)-1-(hexahydrocyclopenta[c]pyrrol-2(1H)-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;-   3-fluoro-N—((R)-1-((S)-2-methylpyrrolidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;    and    or a pharmaceutically acceptable salt thereof.

Methods for obtaining the compounds, or pharmaceutically acceptablesalts thereof,

-   described herein will be apparent to those of ordinary skill in the    art, suitable procedures being described, for example, in examples    below, and in the references cited herein.

Also provided is a method for inhibiting at least one histonedeacetylase. Also provided is a use of at least one compound, orpharmaceutically acceptable salt thereof, described herein in themanufacture of medicament for inhibiting at least one histonedeacetylase. Also provided is at least one compound, or pharmaceuticallyacceptable salt thereof, described herein for use in a method forinhibiting at least one histone deacetylase. In some embodiments, the atleast one histone deacetylase is a Class IIa HDAC. In some embodiments,the at least one histone deacetylase has homology to yeast HDA1. In someembodiments, the at least one histone deacetylase is selected fromHDAC-4, HDAC-5, HDAC-7, and HDAC-9. In some embodiments, the inhibitionis in a cell. In some embodiments, the compound, or pharmaceuticallyacceptable salt thereof, described herein is selective for inhibiting atleast one class II histone deacetylase. In some embodiments, thecompound, or pharmaceutically acceptable salt thereof, described hereinis a selective inhibitor of HDAC-4 and/or HDAC-5.

Also provided is a method of treating a condition or disorder mediatedby HDAC in a subject in need of such a treatment, comprisingadministering to the subject a therapeutically effective amount of atleast one compound, or pharmaceutically acceptable salt thereof,described herein. Also provided is a method of treating a condition ordisorder mediated by HDAC in a subject in need of such a treatment,comprising administering to the subject a therapeutically effectiveamount of at least one compound, or pharmaceutically acceptable saltthereof, described herein. Also provided is a use of at least onecompound, or pharmaceutically acceptable salt thereof, described hereinin the manufacture of medicament for the treatment of a condition ordisorder mediated by HDAC. Also provided is at least one compound, orpharmaceutically acceptable salt thereof, described herein for use in amethod for the treatment of the human or animal body by therapy. Alsoprovided is at least one compound, or pharmaceutically acceptable saltthereof, described herein for use in a method for the treatment of acondition or disorder.

Also provided is a method of treating a condition or disorder responsiveto inhibition of at least one histone deacetylase in a patient in needthereof comprising administering to the patient a therapeuticallyeffective amount of a compound, or pharmaceutically acceptable saltthereof, described herein. In some embodiments, the at least one histonedeacetylase is HDAC-4. In some embodiments, the condition or disorderinvolves a neurodegenerative pathology. In some embodiments, thecondition or disorder is Huntington's disease.

In some embodiments, the condition or disorder mediated by HDACcomprises a neurodegenerative pathology. Accordingly, also provided is amethod of treating a neurodegenerative pathology mediated by HDAC in asubject in need of such a treatment, comprising administering to thesubject a therapeutically effective amount of at least one compound, orpharmaceutically acceptable salt thereof, described herein.

In some embodiments, the neurodegenerative pathology is chosen fromAlzheimer's disease, Parkinson's disease, neuronal intranuclearinclusion disease (NIID), dentatorubral pallidolusyian atrophy (DRPLA),Friedreich's ataxia, Rubenstein-Taubi syndrome, and polyglutaminediseases such as Huntington's disease; spinocerebellar ataxia 1 (SCA 1),spinocerebellar ataxia 7 (SCA 7), seizures, striatonigral degeneration,progressive supranuclear palsy, torsion dystonia, spasmodic torticollis,dyskinesis, familial tremor, Gilles de la Tourette syndrome, diffuseLewy body disease, progressive supranuclear palsy, Pick's disease,primary lateral sclerosis, progressive neural muscular atrophy, spinalmuscular atrophy, hypertrophic interstitial polyneuropathy, retinitispigmentosa, hereditary optic atrophy, hereditary spastic paraplegia,Shy-Drager syndrome, Kennedy's disease, protein-aggregation-relatedneurodegeneration, Machado-Joseph's disease, spongiform encephalopathy,prion-related disease, multiple sclerosis (MS), progressive supranuclearpalsy (Steel-Richardson-Olszewski disease), Hallervorden-Spatz disease,progressive familial myoclonic epilepsy, cerebellar degeneration, motorneuron disease, Werdnig-Hoffman disease, Wohlfart-Kugelberg-Welanderdisease, Charcot-Marie-Tooth disease, Dejerine-Sottas disease, retinitispigmentosa, Leber's disease, progressive systemic sclerosis,dermatomyositis, and mixed connective tissue disease.

In some embodiments, the neurodegenerative pathology is an acute orchronic degenerative disease of the eye. Acute or chronic degenerativediseases of the eye include glaucoma, dry age-related maculardegeneration, retinitis pigmentosa and other forms of heredodegenerativeretinal disease, retinal detachment, macular pucker, ischemia affectingthe outer retina, cellular damage associated with diabetic retinopathyand retinal ischemia, damage associated with laser therapy, ocularneovascular, diabetic retinopathy, rubeosis iritis, uveitis, Fuch'sheterochromatic iridocyclitis, neovascular glaucoma, cornealneovascularization, retinal ischemia, choroidal vascular insufficinency,choroidal thrombosis, carotid artery ischemia, contusive ocular injury,retinopathy of permaturity, retinal vein occlusion, proliferativevitreoretinopathy, corneal angiogenesis, retinal microvasculopathy, andretinal edema.

In some embodiments, the condition or disorder mediated by HDACcomprises a fibrotic disease such as liver fibrosis, cystic fibrosis,cirrhosis, and fibrotic skin diseases, e.g., hypertrophic scars, keloid,and Dupuytren's contracture. Accordingly, also provided is a method oftreating a fibrotic disease mediated by HDAC in a subject in need ofsuch a treatment, comprising administering to the subject atherapeutically effective amount of at least one compound, orpharmaceutically acceptable salt thereof, described herein.

In some embodiments, the condition or disorder mediated by HDACcomprises a psychological disorder, such as depression, bipolar diseaseand dementia. In some embodiments, the condition or disorder mediated byHDAC comprises depression. Accordingly, also provided is a method oftreating a psychological disorder, such as depression, mediated by HDACin a subject in need of such a treatment, comprising administering tothe subject a therapeutically effective amount of at least one compound,or pharmaceutically acceptable salt thereof, described herein. In someembodiments, the depression is chosen from major depressive disorder,and bipolar disorder.

In some embodiments, the condition or disorder mediated by HDACcomprises anxiety. Accordingly, also provided is a method of treating ananxiety mediated by HDAC in a subject in need of such a treatment,comprising administering to the subject a therapeutically effectiveamount of at least one compound, or pharmaceutically acceptable saltthereof, described herein.

In some embodiments, the condition or disorder mediated by HDACcomprises schizophrenia. Accordingly, also provided is a method oftreating a schizophrenia mediated by HDAC in a subject in need of such atreatment, comprising administering to the subject a therapeuticallyeffective amount of at least one compound, or pharmaceuticallyacceptable salt thereof, described herein.

In some embodiments, the condition or disorder mediated by HDACcomprises a motor neuron disease, muscle atrophy/muscle wastingdisorders, or amyotrophic lateral sclerosis (ALS). Accordingly, alsoprovided is a method of treating a motor neuron disease, muscleatrophy/muscle wasting disorders, or amyotrophic lateral sclerosis (ALS)mediated by HDAC in a subject in need of such a treatment, comprisingadministering to the subject a therapeutically effective amount of atleast one compound, or pharmaceutically acceptable salt thereof,described herein.

In some embodiments, the condition or disorder mediated by HDACcomprises a cardiovascular condition. Accordingly, also provided is amethod of treating a cardiovascular condition mediated by HDAC in asubject in need of such a treatment, comprising administering to thesubject a therapeutically effective amount of at least one compound, orpharmaceutically acceptable salt thereof, described herein. In someembodiments, the cardiovascular condition is chosen from cardiomyopathy,cardiac hypertrophy, myocardial ischemia, heart failure, cardiacrestenosis, and arteriosclerosis.

In some embodiments, the condition or disorder mediated by HDACcomprises cancer. Accordingly, also provided is a method of treatingcancer mediated by HDAC in a subject in need of such a treatment,comprising administering to the subject a therapeutically effectiveamount of at least one compound, or pharmaceutically acceptable saltthereof, described herein. In some embodiments, the cancer is chosenfrom lymphoma, pancreatic cancer, colorectal cancer, hepatocellularcarcinoma, Waldenstrom macroglobulinemia, hormone refractory cancer ofthe prostate, and leukaemia, breast cancer, lung cancer, ovarian cancer,prostate cancer, head and neck cancer, renal cancer, gastric cancer,brain cancer, B-cell lymphoma, peripheral T-cell lymphoma, and cutaneousT-cell lymphoma. In some further embodiments, the cancer is chosen fromthe following cancer types. Cardiac: sarcoma (angiosarcoma,fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma,fibroma, lipoma and teratoma; Lung: bronchogenic carcinoma (squamouscell, undifferentiated small cell, undifferentiated large cell,adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma,sarcoma, lymphoma, chondromatous hamartoma, mesothelioma;Gastrointestinal: esophagus (squamous cell carcinoma, adenocarcinoma,leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma,leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma,glucagonoma, gastrinoma, carcinoid tumors, vipoma), small bowel(adenocarcinoma, lymphoma, carcinoid tumors, Karposi's sarcoma,leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large bowel(adenocarcinoma, tubular adenoma, villous adenoma, hamartoma,leiomyoma); Genitourinary tract: kidney (adenocarcinoma, Wilm's tumor[nephroblastoma], lymphoma, leukemia), bladder and urethra (squamouscell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate(adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonalcarcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cellcarcinoma, fibroma, fibroadenoma, adenomatoid tumors, lipoma); Liver:hepatoma, cholangiocarcinoma, hepatoblastoma, angiosarcoma,hepatocellular adenoma, hemangioma; Bone: osteogenic sarcoma(osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma,chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cellsarcoma), multiple myeloma, malignant giant cell tumor chordoma,osteochronfroma (osteocartilaginous exostoses), benign chondroma,chondroblastoma, chondromyxofibroma, osteoid osteoma and giant celltumors; Nervous system: skull (osteoma, hemangioma, granuloma, xanthoma,osteitis deformans), meninges (meningioma, meningiosarcoma,gliomatosis), brain (astrocytoma, medulloblastoma, glioma, ependymoma,germinoma [pinealoma], glioblastoma multiform, oligodendroglioma,schwannoma, retinoblastoma, congenital tumors), spinal cordneurofibroma, meningioma, glioma, sarcoma); Gynecological: uterus(endometrial carcinoma), cervix (cervical carcinoma, pre-tumor cervicaldysplasia), ovaries (ovarian carcinoma [serous cystadenocarcinoma,mucinous cystadenocarcinoma, unclassified carcinoma], granulosa-thecalcell tumors, Sertoli-Leydig cell tumors, dysgerminoma, malignantteratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma,adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma,squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma),fallopian tubes (carcinoma); Hematologic: blood (myeloid leukemia [acuteand chronic], acute lymphoblastic leukemia, chronic lymphocyticleukemia, myeloproliferative diseases, multiple myeloma, myelodysplasticsyndrome), Hodgkin's disease, non-Hodgkin's lymphoma [malignantlymphoma]; Skin: malignant melanoma, basal cell carcinoma, squamous cellcarcinoma, Karposi's sarcoma, moles dysplastic nevi, lipoma, angioma,dermatofibroma, keloids, psoriasis; and Adrenal glands: neuroblastoma.Also provided are methods of sensitization of tumors to radiotherapy byadministering the compound according to the present disclosure before,during or after irradiation of the tumor for treating cancer.

In some embodiments, the condition or disorder mediated by HDACcomprises a condition or disorder treatable by immune modulation.Accordingly, also provided is a method of treating a condition ordisorder treatable by immune modulation mediated by HDAC in a subject inneed of such a treatment, comprising administering to the subject atherapeutically effective amount of at least one compound, orpharmaceutically acceptable salt thereof, described herein. In someembodiments, the condition or disorder treatable by immune modulation ischosen from asthma, irritable bowel syndrome, Crohn's disease,ulcerative colitis, bowel motility disorders, hypertension, rheumatoidarthritis, osteoarthritis, juvenile chronic arthritis, graft versus hostdisease, psoriasis, spondyloarthropathy, inflammatory bowel disease,alcoholic hepatitis, Sjogren's syndrome, ankylosing spondylitis,membranous glomerulopathy, discogenic pain, systemic lupuserythematosus, allergic bowel disease, coeliac disease, bronchitis,cystic fibrosis, rheumatoid spondylitis, osteoarthritis, uveitis,iritis, and conjunctivitis, ischemic bowel disease, psoriasis, eczema,dermatitis, septic arthritis, gout, pseudogout, juvenile arthritis,Still's disease, Henoch-Schonlein purpura, psoriatic arthritis, myalgia,reactive arthritis (Reiter's syndrome), hemochromatosis, Wegener'sgranulomatosis, familial Mediterranean fever (FMF), HBDS(hyperimmunoglobulinemia D and periodic fever syndrome), TRAPS(TNF-alpha receptor associated periodic fever syndrome), chronicobstructive pulmonary disease, neonatal-onset multisystem inflammatorydisease (NOMID), cryopyrin-associated periodic syndrome (CAPS), andfamilial cold autoinflammatory syndrome (FCAS).

In some embodiments, the condition or disorder mediated by HDACcomprises an allergic disease. Accordingly, also provided is a method oftreating an allergic disease, mediated by HDAC in a subject in need ofsuch a treatment, comprising administering to the subject atherapeutically effective amount of at least one compound, orpharmaceutically acceptable salt thereof, described herein. Allergicdiseases include, but are not limited to, respiratory allergic diseasessuch as allergic rhinitis, hypersensitivity lung diseases,hypersensitivity pneumonitis, eosinophilic pneumonias, Loeffler'ssyndrome, chronic eosinophilic pneumonia, delayed-type hypersensitivity,interstitial lung diseases (ILD), idiopathic pulmonary fibrosis,polymyositis, dermatomyositis, systemic anaphylaxis, drug allergies(e.g., to penicillin or cephalosporins), and insect sting allergies.

In some embodiments, the condition or disorder mediated by HDACcomprises an infectious disease such as a fungal infection, bacterialinfection, viral infection, and protozoal infection, e.g., malaria,giardiasis, leishmaniasis, Chaga's disease, dysentery, toxoplasmosis,and coccidiosis. In some embodiments, the condition or disorder mediatedby HDAC comprises malaria. Accordingly, also provided is a method oftreating an infectious disease, such as malaria, mediated by HDAC in asubject in need of such a treatment, comprising administering to thesubject a therapeutically effective amount of at least one compound, orpharmaceutically acceptable salt thereof, described herein.

In some embodiments, the condition or disorder mediated by HDACcomprises autism or Rett syndrome. Accordingly, also provided is amethod of treating autism or Rett syndrome mediated by HDAC in a subjectin need of such a treatment, comprising administering to the subject atherapeutically effective amount of at least one compound, orpharmaceutically acceptable salt thereof, described herein.

In some embodiments, the condition or disorder mediated by HDACcomprises a hematological disorder such as thalassemia, anemia, andsickle cell anemia. Accordingly, also provided is a method of treating ahematological disorder mediated by HDAC in a subject in need of such atreatment, comprising administering to the subject a therapeuticallyeffective amount of at least one compound, or pharmaceuticallyacceptable salt thereof, described herein.

In some embodiments, the condition or disorder mediated by HDACcomprises a metabolic disease such as prediabetes or diabetes (type I orII). Accordingly, also provided is a method of treating a metabolicdisease, such as prediabetes or diabetes (type I or II), mediated byHDAC in a subject in need of such a treatment, comprising administeringto the subject a therapeutically effective amount of at least onecompound, or pharmaceutically acceptable salt thereof, described herein.

In some embodiments, the condition or disorder mediated by HDACcomprises a disorder that may also be treated by progenitor/stem cellbased therapies such as: disorders related to diabetes (organ failure,cirrhosis, and hepatitis); central nervous system (CNS) disordersassociated with dysregulation of progenitor cells in the brain (e.g.,post-traumatic stress disorder (PTSD); tumors (e.g., retinoblastomas);disorders affecting oligodendrocyte progenitor cells (e.g., astrocytomasand ependimal cell tumors); multiple sclerosis; demyelinating disorderssuch as the leukodystrophies; neuropathies associated with white matterloss; and cerebellar disorders such as ataxia; and olfactory progenitordisorders (e.g., anosmic conditions). Accordingly, also provided is amethod of treating a disorder that is mediated by HDAC in a subject inneed of such a treatment, comprising administering to the subject atherapeutically effective amount of at least one compound, orpharmaceutically acceptable salt thereof, described herein, eitherbefore, during, or after a treatment with progenitor/stem cell basedtherapies.

In some embodiments, the condition or disorder mediated by HDACcomprises a disorder related to the proliferation of epithelial andmesenchymal cells (e.g., tumors, wound healing, and surgeries).Accordingly, also provided is a method of treating a disorder related tothe proliferation of epithelial and mesenchymal cells that is mediatedby HDAC in a subject in need of such a treatment, comprisingadministering to the subject a therapeutically effective amount of atleast one compound, or pharmaceutically acceptable salt thereof,described herein.

In some embodiments, the condition or disorder mediated by HDACcomprises a disorder related to the proliferation of bone progenitors(e.g., osteoblasts and osteoclasts), disorders related to hair andepidermal progenitors (e.g., hair loss, cutaneous tumors, skinregeneration, burns, and cosmetic surgery); and disorders related tobone loss during menopause. Accordingly, also provided is a method oftreating disorders related to the proliferation of bone progenitors,disorders related to hair and epidermal progenitors, or disordersrelated to bone loss that are mediated by HDAC in a subject in need ofsuch a treatment, comprising administering to the subject atherapeutically effective amount of at least one compound, orpharmaceutically acceptable salt thereof, described herein.

In some embodiments, the condition or disorder mediated by HDAC is aviral disorder for which blood cells become sensitized to othertreatments after HDAC inhibition, following administering to the subjecta therapeutically effective amount of at least one compound, orpharmaceutically acceptable salt thereof, as described herein.Accordingly, also provided is a method of treating a viral disorder,wherein blood cells become sensitized to other treatments after HDACinhibition, that is mediated by HDAC in a subject in need of such atreatment, comprising administering to the subject a therapeuticallyeffective amount of at least one compound, or pharmaceuticallyacceptable salt thereof, described herein.

In some embodiments, the condition or disorder mediated by HDAC is animmune disorder that may be co-treated with TNFα or other immunemodulators, upon administering to the subject a therapeuticallyeffective amount of at least one compound, or pharmaceuticallyacceptable salt thereof, as described herein. Accordingly, also providedis a method of treating an immune disorder that is mediated by HDAC in asubject in need of such a treatment, comprising administering to thesubject a therapeutically effective amount of at least one compound, orpharmaceutically acceptable salt thereof, described herein, eitherbefore, during, or after a treatment with TNFα or other immunemodulators.

In some embodiments, the condition or disorder mediated by HDACcomprises a graft rejection or transplant rejection. Accordingly, alsoprovided is a method of treating a disorder related to a graft rejectionor a transplant rejection that is mediated by HDAC in a subject in needof such a treatment, comprising administering to the subject atherapeutically effective amount of at least one compound, orpharmaceutically acceptable salt thereof, described herein.

In some embodiments, the condition or disorder mediated by HDACcomprises a blood pressure disorder related to nitric oxide (NO)regulation (e.g., hypertension, erectile dysfunction, asthma; and oculardisorders as glaucoma). Accordingly, also provided is a method oftreating a blood pressure disorder related to nitric oxide (NO)regulation that is mediated by HDAC in a subject in need of such atreatment, comprising administering to the subject a therapeuticallyeffective amount of at least one compound, or pharmaceuticallyacceptable salt thereof, described herein. In some embodiments, thecondition or disorder is a cardiac hypertrophic disorder. Accordingly,also provided is a method of treating a cardiac hypertrophic disorderthat is mediated by HDAC in a subject in need of such a treatment,comprising administering to the subject a therapeutically effectiveamount of at least one compound, or pharmaceutically acceptable saltthereof, described herein.

Also provided are methods of treatment in which at least one compound,or pharmaceutically acceptable salt thereof, described herein is theonly active agent given to the subject and methods of treatment in whichat least one compound, or pharmaceutically acceptable salt thereof,described herein is given to the subject in combination with one or moreadditional active agents.

In general, the compounds, or pharmaceutically acceptable salts thereof,described herein will be administered in a therapeutically effectiveamount by any of the accepted modes of administration for agents thatserve similar utilities. The actual amount of the compound, i.e., theactive ingredient, will depend upon numerous factors such as theseverity of the disease to be treated, the age and relative health ofthe subject, the potency of the compound used, the route and form ofadministration, and other factors well known to the skilled artisan. Thedrug can be administered at least once a day, such as once or twice aday.

In some embodiments, the compounds, or pharmaceutically acceptable saltsthereof, described herein are administered as a pharmaceuticalcomposition. Accordingly, provided are pharmaceutical compositionscomprising at least one compound, or pharmaceutically acceptable saltthereof, described herein, together with at least one pharmaceuticallyacceptable vehicle chosen from carriers, adjuvants, and excipients. Acompound of the present disclosure can be formulated into pharmaceuticalcompositions using techniques well known to those in the art.

Pharmaceutically acceptable vehicles must be of sufficiently high purityand sufficiently low toxicity to render them suitable for administrationto the animal being treated. The vehicle can be inert or it can possesspharmaceutical benefits. The amount of vehicle employed in conjunctionwith the compound, or pharmaceutically acceptable salt thereof, issufficient to provide a practical quantity of material foradministration per unit dose of the compound, or pharmaceuticallyacceptable salt thereof.

Exemplary pharmaceutically acceptable carriers or components thereof aresugars, such as lactose, glucose and sucrose; starches, such as cornstarch and potato starch; cellulose and its derivatives, such as sodiumcarboxymethyl cellulose, ethyl cellulose, and methyl cellulose; powderedtragacanth; malt; gelatin; talc; solid lubricants, such as stearic acidand magnesium stearate; calcium sulfate; synthetic oils; vegetable oils,such as peanut oil, cottonseed oil, sesame oil, olive oil, and corn oil;polyols such as propylene glycol, glycerine, sorbitol, mannitol, andpolyethylene glycol; alginic acid; phosphate buffer solutions;emulsifiers, such as the TWEENs®; wetting agents, such sodium laurylsulfate; coloring agents; flavoring agents; tableting agents;stabilizers; antioxidants; preservatives; pyrogen-free water; isotonicsaline; and phosphate buffer solutions.

Optional active agents may be included in a pharmaceutical composition,which do not substantially interfere with the activity of the compound,or pharmaceutically acceptable salt thereof, described herein.

Effective concentrations of at least one compound, or pharmaceuticallyacceptable salt thereof, described herein are mixed with a suitablepharmaceutically acceptable vehicle. In instances in which the compound,or pharmaceutically acceptable salt thereof, exhibits insufficientsolubility, methods for solubilizing compounds may be used. Such methodsare known to those of skill in this art, and include, but are notlimited to, using cosolvents, such as dimethylsulfoxide (DMSO), usingsurfactants, such as TWEEN®, or dissolution in aqueous sodiumbicarbonate.

Upon mixing or addition of a compound, or pharmaceutically acceptablesalt thereof, described herein, the resulting mixture may be a solution,suspension, emulsion or the like. The form of the resulting mixturedepends upon a number of factors, including the intended mode ofadministration and the solubility of the compound, or pharmaceuticallyacceptable salt thereof, in the chosen vehicle. The effectiveconcentration sufficient for ameliorating the symptoms of the diseasetreated may be empirically determined.

The compounds, or pharmaceutically acceptable salts thereof, describedherein may be administered orally, topically, parenterally,intravenously, by intramuscular injection, by inhalation or spray,sublingually, transdermally, via buccal administration, rectally, as anophthalmic solution, or by other means, in dosage unit formulations.

Pharmaceutical compositions may be formulated for oral use, such as forexample, tablets, troches, lozenges, aqueous or oily suspensions,dispersible powders or granules, emulsions, hard or soft capsules, orsyrups or elixirs. Pharmaceutical compositions intended for oral use maybe prepared according to any method known to the art for the manufactureof pharmaceutical compositions and such compositions may contain one ormore agents, such as sweetening agents, flavoring agents, coloringagents and preserving agents, in order to provide pharmaceuticallyelegant and palatable preparations. In some embodiments, oralpharmaceutical compositions contain from 0.1 to 99% of at least onecompound, or pharmaceutically acceptable salt thereof, described herein.In some embodiments, oral pharmaceutical compositions contain at least5% (weight %) of at least one compound, or pharmaceutically acceptablesalt thereof, described herein. Some embodiments contain from 25% to 50%or from 5% to 75% of at least one compound, or pharmaceuticallyacceptable salt thereof, described herein.

Orally administered pharmaceutical compositions also include liquidsolutions, emulsions, suspensions, powders, granules, elixirs,tinctures, syrups, and the like. The pharmaceutically acceptablecarriers suitable for preparation of such compositions are well known inthe art. Oral pharmaceutical compositions may contain preservatives,flavoring agents, sweetening agents, such as sucrose or saccharin,taste-masking agents, and coloring agents.

Typical components of carriers for syrups, elixirs, emulsions andsuspensions include ethanol, glycerol, propylene glycol, polyethyleneglycol, liquid sucrose, sorbitol and water. Syrups and elixirs may beformulated with sweetening agents, for example glycerol, propyleneglycol, sorbitol or sucrose. Such pharmaceutical compositions may alsocontain a demulcent.

The compound, or pharmaceutically acceptable salt thereof, describedherein can be incorporated into oral liquid preparations such as aqueousor oily suspensions, solutions, emulsions, syrups, or elixirs, forexample. Furthermore, pharmaceutical compositions containing thecompound, or pharmaceutically acceptable salt thereof, described hereincan be presented as a dry product for constitution with water or othersuitable vehicle before use. Such liquid preparations can containconventional additives, such as suspending agents (e.g., sorbitol syrup,methyl cellulose, glucose/sugar, syrup, gelatin, hydroxyethyl cellulose,carboxymethyl cellulose, aluminum stearate gel, and hydrogenated ediblefats), emulsifying agents (e.g., lecithin, sorbitan monooleate, oracacia), non-aqueous vehicles, which can include edible oils (e.g.,almond oil, fractionated coconut oil, silyl esters, propylene glycol andethyl alcohol), and preservatives (e.g., methyl or propylp-hydroxybenzoate and sorbic acid).

For a suspension, typical suspending agents include methylcellulose,sodium carboxymethyl cellulose, Avicel® RC-591, tragacanth and sodiumalginate; typical wetting agents include lecithin and polysorbate 80;and typical preservatives include methyl paraben and sodium benzoate.

Aqueous suspensions contain the active material(s) in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydropropylmethylcellulose,sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents; may be a naturally-occurring phosphatide,for example, lecithin, or condensation products of an alkylene oxidewith fatty acids, for example polyoxyethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethyleneoxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol substitute, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan substitute.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, or n-propyl p-hydroxybenzoate.

Oily suspensions may be formulated by suspending the active ingredientsin a vegetable oil, for example peanut oil, olive oil, sesame oil orcoconut oil, or in a mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide palatable oralpreparations. These pharmaceutical compositions may be preserved by theaddition of an anti-oxidant such as ascorbic acid.

Pharmaceutical compositions may also be in the form of oil-in-wateremulsions. The oily phase may be a vegetable oil, for example olive oilor peanut oil, or a mineral oil, for example liquid paraffin or mixturesof these. Suitable emulsifying agents may be naturally-occurring gums,for example gum acacia or gum tragacanth, naturally-occurringphosphatides, for example soy bean, lecithin, and esters or partialesters derived from fatty acids and hexitol, anhydrides, for examplesorbitan monooleate, and condensation products of the said partialesters with ethylene oxide, for example polyoxyethylene sorbitanmonooleate.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.

Tablets typically comprise conventional pharmaceutically acceptableadjuvants as inert diluents, such as calcium carbonate, sodiumcarbonate, mannitol, lactose and cellulose; binders such as starch,gelatin and sucrose; disintegrants such as starch, alginic acid andcroscarmellose; lubricants such as magnesium stearate, stearic acid andtalc. Glidants such as silicon dioxide can be used to improve flowcharacteristics of the powder mixture. Coloring agents, such as the FD&Cdyes, can be added for appearance. Sweeteners and flavoring agents, suchas aspartame, saccharin, menthol, peppermint, and fruit flavors, can beuseful adjuvants for chewable tablets. Capsules (including time releaseand sustained release formulations) typically comprise one or more soliddiluents disclosed above. The selection of carrier components oftendepends on secondary considerations like taste, cost, and shelfstability.

Such pharmaceutical compositions may also be coated by conventionalmethods, typically with pH or time-dependent coatings, such that thecompound, or pharmaceutically acceptable salt thereof, is released inthe gastrointestinal tract in the vicinity of the desired topicalapplication, or at various times to extend the desired action. Suchdosage forms typically include, but are not limited to, one or more ofcellulose acetate phthalate, polyvinylacetate phthalate, hydroxypropylmethylcellulose phthalate, ethyl cellulose, Eudragit® coatings, waxesand shellac.

Pharmaceutical compositions for oral use may also be presented as hardgelatin capsules wherein the active ingredient is mixed with an inertsolid diluent, for example, calcium carbonate, calcium phosphate orkaolin, or as soft gelatin capsules wherein the active ingredient ismixed with water or an oil medium, for example peanut oil, liquidparaffin or olive oil.

Pharmaceutical compositions may be in the form of a sterile injectableaqueous or oleaginous suspension. This suspension may be formulatedaccording to the known art using those suitable dispersing or wettingagents and suspending agents that have been mentioned above. The sterileinjectable preparation may also be sterile injectable solution orsuspension in a non-toxic parentally acceptable vehicle, for example asa solution in 1,3-butanediol. Among the acceptable vehicles that may beemployed are water, Ringer's solution, and isotonic sodium chloridesolution. In addition, sterile, fixed oils are conventionally employedas a solvent or suspending medium. For this purpose any bland fixed oilmay be employed including synthetic mono- or diglycerides. In addition,fatty acids such as oleic acid can be useful in the preparation ofinjectables.

The compound, or pharmaceutically acceptable salt thereof, describedherein may be administered parenterally in a sterile medium. Parenteraladministration includes subcutaneous injections, intravenous,intramuscular, intrathecal injection or infusion techniques. Thecompound, or pharmaceutically acceptable salt thereof, described herein,depending on the vehicle and concentration used, can either be suspendedor dissolved in the vehicle. Advantageously, adjuvants such as localanesthetics, preservatives and buffering agents can be dissolved in thevehicle. In many pharmaceutical compositions for parenteraladministration the carrier comprises at least 90% by weight of the totalcomposition. In some embodiments, the carrier for parenteraladministration is chosen from propylene glycol, ethyl oleate,pyrrolidone, ethanol, and sesame oil.

The compound, or pharmaceutically acceptable salt thereof, describedherein may also be administered in the form of suppositories for rectaladministration of the drug. These pharmaceutical compositions can beprepared by mixing the drug with a suitable non-irritating excipientthat is solid at ordinary temperatures but liquid at rectal temperatureand will therefore melt in the rectum to release the drug. Suchmaterials include cocoa butter and polyethylene glycols.

The compound, or pharmaceutically acceptable salt thereof, describedherein may be formulated for local or topical application, such as fortopical application to the skin and mucous membranes, such as in theeye, in the form of gels, creams, and lotions and for application to theeye. Topical pharmaceutical compositions may be in any form including,for example, solutions, creams, ointments, gels, lotions, milks,cleansers, moisturizers, sprays, skin patches, and the like.

Such solutions may be formulated as 0.01%-10% isotonic solutions, pH5-7, with appropriate salts. The compound, or pharmaceuticallyacceptable salt thereof, described herein may also be formulated fortransdermal administration as a transdermal patch.

Topical pharmaceutical compositions comprising at least one compound, orpharmaceutically acceptable salt thereof, described herein can beadmixed with a variety of carrier materials well known in the art, suchas, for example, water, alcohols, aloe vera gel, allantoin, glycerine,vitamin A and E oils, mineral oil, propylene glycol, PPG-2 myristylpropionate, and the like.

Other materials suitable for use in topical carriers include, forexample, emollients, solvents, humectants, thickeners and powders.Examples of each of these types of materials, which can be used singlyor as mixtures of one or more materials, are as follows.

Representative emollients include stearyl alcohol, glycerylmonoricinoleate, glyceryl monostearate, propane-1,2-diol,butane-1,3-diol, mink oil, cetyl alcohol, iso-propyl isostearate,stearic acid, iso-butyl palmitate, isocetyl stearate, oleyl alcohol,isopropyl laurate, hexyl laurate, decyl oleate, octadecan-2-ol, isocetylalcohol, cetyl palmitate, dimethylpolysiloxane, di-n-butyl sebacate,iso-propyl myristate, iso-propyl palmitate, iso-propyl stearate, butylstearate, polyethylene glycol, triethylene glycol, lanolin, sesame oil,coconut oil, arachis oil, castor oil, acetylated lanolin alcohols,petroleum, mineral oil, butyl myristate, isostearic acid, palmitic acid,isopropyl linoleate, lauryl lactate, myristyl lactate, decyl oleate, andmyristyl myristate; propellants, such as propane, butane, iso-butane,dimethyl ether, carbon dioxide, and nitrous oxide; solvents, such asethyl alcohol, methylene chloride, iso-propanol, castor oil, ethyleneglycol monoethyl ether, diethylene glycol monobutyl ether, diethyleneglycol monoethyl ether, dimethyl sulphoxide, dimethyl formamide,tetrahydrofuran; humectants, such as glycerin, sorbitol, sodium2-pyrrolidone-5-carboxylate, soluble collagen, dibutyl phthalate, andgelatin; and powders, such as chalk, talc, fullers earth, kaolin,starch, gums, colloidal silicon dioxide, sodium polyacrylate, tetraalkyl ammonium smectites, trialkyl aryl ammonium smectites, chemicallymodified magnesium aluminium silicate, organically modifiedmontmorillonite clay, hydrated aluminium silicate, fumed silica,carboxyvinyl polymer, sodium carboxymethyl cellulose, and ethyleneglycol monostearate.

The compound, or pharmaceutically acceptable salt thereof, describedherein may also be topically administered in the form of liposomedelivery systems, such as small unilamellar vesicles, large unilamellarvesicles, and multilamellar vesicles. Liposomes can be formed from avariety of phospholipids, such as cholesterol, stearylamine orphosphatidylcholines.

Other pharmaceutical compositions useful for attaining systemic deliveryof the compound, or pharmaceutically acceptable salt thereof, includesublingual, buccal and nasal dosage forms. Such pharmaceuticalcompositions typically comprise one or more of soluble filler substancessuch as sucrose, sorbitol and mannitol, and binders such as acacia,microcrystalline cellulose, carboxymethyl cellulose, and hydroxypropylmethylcellulose. Glidants, lubricants, sweeteners, colorants,antioxidants and flavoring agents disclosed above may also be included.

Pharmaceutical compositions for inhalation typically can be provided inthe form of a solution, suspension or emulsion that can be administeredas a dry powder or in the form of an aerosol using a conventionalpropellant (e.g., dichlorodifluoromethane or trichlorofluoromethane).

The pharmaceutical compositions may also optionally comprise an activityenhancer. The activity enhancer can be chosen from a wide variety ofmolecules that function in different ways to enhance or be independentof therapeutic effects of the compound, or pharmaceutically acceptablesalt thereof, described herein. Particular classes of activity enhancersinclude skin penetration enhancers and absorption enhancers.

Pharmaceutical compositions may also contain additional active agentsthat can be chosen from a wide variety of molecules, which can functionin different ways to enhance the therapeutic effects of at least onecompound, or pharmaceutically acceptable salt thereof, described herein.These optional other active agents, when present, are typically employedin the pharmaceutical compositions at a level ranging from 0.01% to 15%.Some embodiments contain from 0.1% to 10% by weight of the composition.Other embodiments contain from 0.5% to 5% by weight of the composition.

Also provided are packaged pharmaceutical compositions. Such packagedcompositions include a pharmaceutical composition comprising at leastone compound, or pharmaceutically acceptable salt thereof, describedherein, and instructions for using the composition to treat a subject(typically a human patient). In some embodiments, the instructions arefor using the pharmaceutical composition to treat a subject suffering acondition or disorder mediated by HDAC. The packaged pharmaceuticalcomposition can include providing prescribing information; for example,to a patient or health care provider, or as a label in a packagedpharmaceutical composition. Prescribing information may include forexample efficacy, dosage and administration, contraindication andadverse reaction information pertaining to the pharmaceuticalcomposition.

In all of the foregoing the compound, or pharmaceutically acceptablesalt thereof, can be administered alone, as mixtures, or in combinationwith other active agents.

The methods described herein include methods for treating Huntington'sdisease, including treating memory and/or cognitive impairmentassociated with Huntington's disease, comprising administering to asubject, simultaneously or sequentially, at least one compound, orpharmaceutically acceptable salt thereof, described herein and one ormore additional agents used in the treatment of Huntington's diseasesuch as, but not limited to, Amitriptyline, Imipramine, Desipramine,Nortriptyline, Paroxetine, Fluoxetine, Sertraline, Tetrabenazine,Haloperidol, Chlorpromazine, Thioridazine, Sulpride, Quetiapine,Clozapine, and Risperidone. In methods using simultaneousadministration, the agents can be present in a combined composition orcan be administered separately. As a result, also provided arepharmaceutical compositions comprising at least one compound, orpharmaceutically acceptable salt thereof, described herein and one ormore additional pharmaceutical agents used in the treatment ofHuntington's disease such as, but not limited to, Amitriptyline,Imipramine, Desipramine, Nortriptyline, Paroxetine, Fluoxetine,Setraline, Terabenazine, Haloperidol, Chlorpromazine, Thioridazine,Sulpride, Quetiapine, Clozapine, and Risperidone. Similarly, alsoprovided are packaged pharmaceutical compositions containing apharmaceutical composition comprising at least one compound, orpharmaceutically acceptable salt thereof, described herein, and anothercomposition comprising one or more additional pharmaceutical agents usedin the treatment of Huntington's disease such as, but not limited to,Amitriptyline, Imipramine, Desipramine, Nortriptyline, Paroxetine,Fluoxetine, Setraline, Terabenazine, Haloperidol, Chlorpromazine,Thioridazine, Sulpride, Quetiapine, Clozapine, and Risperidone.

Also provided are methods for treating Alzheimer's disease, includingtreating memory and/or cognitive impairment associated with Alzheimer'sdisease, comprising administering to a subject, simultaneously orsequentially, at least one compound, or pharmaceutically acceptable saltthereof, described herein and one or more additional agents used in thetreatment of Alzheimer's disease such as, but not limited to, Reminyl®,Cognex®, Aricept®, Exelon®, Akatinol®, Neotropin™, Eldepryl®, Estrogenand Clioquinol. In methods using simultaneous administration, the agentscan be present in a combined composition or can be administeredseparately. Also provided are pharmaceutical compositions comprising atleast one compound, or pharmaceutically acceptable salt thereof,described herein, and one or more additional pharmaceutical agents usedin the treatment of Alzheimer's disease such as, but not limited to,Reminyl®, Cognex®, Aricept®, Exelon®, Akatinol®, Neotropin™, Eldepryl®,Estrogen and Clioquinol. Similarly, also provided are packagedpharmaceutical compositions containing a pharmaceutical compositioncomprising at least one compound, or pharmaceutically acceptable saltthereof, described herein, and another composition comprising one ormore additional pharmaceutical agents used in the treatment ofAlzheimer's disease such as, but not limited to Reminyl®, Cognex®,Aricept®, Exelon®, Akatinol®, Neotropin™, Eldepryl®, Estrogen andClioquinol.

Also provided are methods for treating cancer comprising administeringto a subject, simultaneously or sequentially, at least one compound, orpharmaceutically acceptable salt thereof, described herein and one ormore additional agents used in the treatment of cancer such as, but notlimited to, the following categories of anti-tumor agents:

(i) other cell cycle inhibitory agents that work by the same ordifferent mechanisms from those defined herein before, for examplecyclin dependent kinase (CDK) inhibitors, in particular CDK2 inhibitors;

(ii) cytostatic agents such as antioestrogens (for example tamoxifen,toremifene, raloxifene, droloxifene, iodoxyfene), progestogens (forexample megestrol acetate), aromatase inhibitors (for exampleanastrozole, letrazole, vorazole, exemestane), antiprogestogens,antiandrogens (for example flutamide, nilutamide, bicalutamide,cyproterone acetate), LHRH agonists and antagonists (for examplegoserelin acetate, luprolide), inhibitors of testosterone5α-dihydroreductase (for example finasteride), anti-invasion agents (forexample metalloproteinase inhibitors like marimastat and inhibitors ofurokinase plasminogen activator receptor function) and inhibitors ofgrowth factor function, (such growth factors include for examplevascular endothelial growth factor, epithelial growth factor, plateletderived growth factor and hepatocyte growth factor such inhibitorsinclude growth factor antibodies, growth factor receptor antibodies,tyrosine kinase inhibitors and serine/threonine kinase inhibitors);(iii) antiproliferative/antineoplastic drugs and combinations thereof,as used in medical oncology, such as antimetabolites (for exampleantifolates like methotrexate, fluoropyrimidines like 5-fluorouracil,purine and adenosine analogues, cytosine arabinoside); antitumourantibiotics (for example anthracyclines like doxorubicin, daunomycin,epirubicin and idarubicin, mitomycin-C, dactinomycin, mithramycin);platinum derivatives (for example cisplatin, carboplatin); alkylatingagents (for example nitrogen mustard, melphalan, chlorambucil,busulphan, cyclophosphamide, ifosfamide, nitrosoureas, thiotepa);antimitotic agents (for example vinca alkaloids like vincrisitine andtaxoids like taxol, taxotere); topoisomerase inhibitors (for exampleepipodophyllotoxins like etoposide and teniposide, amsacrine,topotecan);(iv) antiangiogenic agents that work by different mechanisms from thosedefined herein before (for example receptor tyrosine kinases like Tie-2,inhibitors of integrin α_(V)β₃ function, angiostatin, razoxin,thalidomide), and including vascular targeting agents; and(v) differentiation agents (for example retinoic acid and vitamin D).

In methods using simultaneous administration, the agents can be presentin a combined composition or can be administered separately. Alsoprovided are pharmaceutical compositions comprising at least onecompound, or pharmaceutically acceptable salt thereof, described herein,and one or more anti-tumor agent as described herein. Similarly, alsoprovided are packaged pharmaceutical compositions containing apharmaceutical composition comprising at least one compound, orpharmaceutically acceptable salt thereof, described herein, and anothercomposition comprising one or more one or more anti-tumor agent asdescribed herein. When used in combination with one or more additionalpharmaceutical agent or agents, the compounds described herein may beadministered prior to, concurrently with, or following administration ofthe additional pharmaceutical agent or agents.

In some embodiments, the compounds, or pharmaceutically acceptable saltsthereof, described herein, are administered in conjunction with surgeryor radiotherapy, optionally in combination with one or more additionalagents used in the treatment of cancer.

The dosages of the compounds described herein depend upon a variety offactors including the particular syndrome to be treated, the severity ofthe symptoms, the route of administration, the frequency of the dosageinterval, the particular compound utilized, the efficacy, toxicologyprofile, pharmacokinetic profile of the compound, and the presence ofany deleterious side-effects, among other considerations.

The compound, or pharmaceutically acceptable salt thereof, describedherein is typically administered at dosage levels and in a mannercustomary for HDAC inhibitors. For example, the compound, orpharmaceutically acceptable salt thereof, can be administered, in singleor multiple doses, by oral administration at a dosage level of generally0.001-100 mg/kg/day, for example, 0.01-100 mg/kg/day, such as 0.1-70mg/kg/day, for example, 0.5-10 mg/kg/day. Unit dosage forms can containgenerally 0.01-1000 mg of at least one compound, or pharmaceuticallyacceptable salt thereof, described herein, for example, 0.1-50 mg of atleast one compound, or pharmaceutically acceptable salt thereof,described herein. For intravenous administration, the compounds can beadministered, in single or multiple dosages, at a dosage level of, forexample, 0.001-50 mg/kg/day, such as 0.001-10 mg/kg/day, for example,0.01-1 mg/kg/day. Unit dosage forms can contain, for example, 0.1-10 mgof at least one compound, or pharmaceutically acceptable salt thereof,described herein.

A labeled form of a compound, or pharmaceutically acceptable saltthereof, described herein can be used as a diagnostic for identifyingand/or obtaining compounds that have the function of modulating anactivity of HDAC as described herein. The compound, or pharmaceuticallyacceptable salt thereof, described herein may additionally be used forvalidating, optimizing, and standardizing bioassays.

By “labeled” herein is meant that the compound is either directly orindirectly labeled with a label which provides a detectable signal,e.g., radioisotope, fluorescent tag, enzyme, antibodies, particles suchas magnetic particles, chemiluminescent tag, or specific bindingmolecules, etc. Specific binding molecules include pairs, such as biotinand streptavidin, digoxin and antidigoxin etc. For the specific bindingmembers, the complementary member would normally be labeled with amolecule which provides for detection, in accordance with knownprocedures, as outlined above. The label can directly or indirectlyprovide a detectable signal.

The present disclosure includes all isotopes of atoms occurring in thecompounds and pharmaceutically acceptable salts thereof describedherein. Isotopes include those atoms having the same atomic number butdifferent mass numbers. The present disclosure also includes everycombination of one or more atoms in the compounds and pharmaceuticallyacceptable salts thereof described herein that is replaced with an atomhaving the same atomic number but a different mass number. One suchexample is the replacement of an atom that is the most naturallyabundant isotope, such as ¹H or ¹²C, found in one of the compounds andpharmaceutically acceptable salts thereof described herein, with adifferent atom that is not the most naturally abundant isotope, such as²H or ³H (replacing ¹H), or ¹¹C, ¹³C, or ¹²C (replacing ¹²C). A compoundwherein such a replacement has taken place is commonly referred to asbeing isotopically-labeled. Isotopic-labeling of the compounds andpharmaceutically acceptable salts thereof described herein can beaccomplished using any one of a variety of different synthetic methodsknow to those of ordinary skill in the art and they are readily creditedwith understanding the synthetic methods and available reagents neededto conduct such isotopic-labeling. By way of general example, andwithout limitation, isotopes of hydrogen include ²H (deuterium) and ³H(tritium). Isotopes of carbon include ¹¹C, ¹³C, and ¹⁴C. Isotopes ofnitrogen include ¹³N and ¹⁵N. Isotopes of oxygen include ¹⁵O, ¹⁷O, and¹⁸O. An isotope of fluorine includes ¹⁸F. An isotope of sulfur includes³⁵S. An isotope of chlorine includes ³⁶Cl. Isotopes of bromine include⁷⁵Br, ⁷⁶Br, ⁷⁷Br, and ⁸²Br. Isotopes of iodine include ¹²³I, ¹²⁴I, ¹²⁵I,and ¹³¹I. Also provided are pharmaceutical compositions comprising acompound or a pharmaceutically acceptable salt thereof described herein,wherein the naturally occurring distribution of the isotopes in thepharmaceutical composition is perturbed. Also provided arepharmaceutical compositions comprising a compound or a pharmaceuticallyacceptable salt thereof described herein enriched at one or morepositions with an isotope other than the most naturally abundantisotope. Methods are readily available to measure such isotopeperturbations or enrichments, such as, mass spectrometry, and forisotopes that are radio-isotopes additional methods are available, suchas, radio-detectors used in connection with HPLC or gas chromatography(GC). Certain isotopically-labeled compounds and pharmaceuticallyacceptable salts thereof described herein are useful in compound and/orsubstrate tissue distribution assays. In some embodiments theradionuclide ³H and/or ¹⁴C isotopes are useful in these studies.Further, substitution with heavier isotopes such as deuterium (i.e., ²H)may afford certain therapeutic advantages resulting from greatermetabolic stability (e.g., increased in vivo half-life or reduced dosagerequirements) and hence may be preferred in some circumstances.Isotopically labeled compounds and pharmaceutically acceptable saltsthereof described herein can generally be prepared by followingprocedures analogous to those disclosed in the Examples infra, bysubstituting an isotopically labeled reagent for a non-isotopicallylabeled reagent. Moreover, it should be understood that all of the atomsrepresented in the compounds and pharmaceutically acceptable saltsthereof described herein can be either the most commonly occurringisotope of such atoms or a scarcer radio-isotope or nonradioactiveisotope.

In carrying out the procedures of the methods described herein, it is ofcourse to be understood that reference to particular buffers, media,reagents, cells, culture conditions and the like are not intended to belimiting, but are to be read so as to include all related materials thatone of ordinary skill in the art would recognize as being of interest orvalue in the particular context in which that discussion is presented.For example, it is often possible to substitute one buffer system orculture medium for another and still achieve similar, if not identical,results. Those of skill in the art will have sufficient knowledge ofsuch systems and methodologies so as to be able, without undueexperimentation, to make such substitutions as will optimally servetheir purposes in using the methods and procedures disclosed herein.

EXAMPLES

The compounds, or pharmaceutically acceptable salts thereof,compositions, and methods described herein are further illustrated bythe following non-limiting examples.

As used herein, the following abbreviations have the following meanings.If an abbreviation is not defined, it has its generally acceptedmeaning.

Abbreviations

-   AcOH Acetic acid-   Bn: Benzyl-   BOC or Boc: tert-butyloxycarbonyl-   cPr: cyclopropyl-   DCM: Dichloromethane-   DIPE: Diisopropylether-   DIPEA: Diisopropylethylamine-   DMAP: Dimethylaminopyridine-   DMF: Dimethylformamide-   DMSO: Dimethylsulfoxide-   EDC: 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide-   eqs: equivalents-   ES+: Electrospray Positive Ionisation-   Et₂O: Diethyl ether-   EtOAc: Ethyl acetate-   EtOH: Ethanol-   FBS Fetal bovine serum-   h: Hour-   HATU:    (1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium    3-oxid hexafluorophosphate)-   HBTU: N,N,N′,N′-Tetramethyl-O-(1H-benzotriazol-1-yl)uronium    hexafluorophosphate-   HOPO: 2-Hydroxypyridine-N-oxide-   HPLC: High Performance Liquid Chromatography-   i-hex: iso-Hexane-   IPA: iso-Propyl alcohol-   LCMS: Liquid Chromatography Mass Spectrometry-   M: Mass-   Me: Methyl-   MeCN: Acetonitrile-   MeOH: Methanol-   min: minute(s)-   Ms: Mesyl-   NMR: Nuclear Magnetic Resonance-   Pd/C: Palladium on carbon-   Ph: Phenyl-   RT: Retention time-   r.t.: Room temperature-   SFC: Supercritical Fluid Chromatography-   SCX: Strong Cation Exchange-   TFA: Trifluoroacetic acid-   TFAA: Trifluoroacetic anhydride-   THF: Tetrahydrofuran-   v/v or V/V: volume to volume

Compounds were named with the aid of the Cambridgesoft ChemistryCartridge (v. 9.0.0.182) software.

All reactions involving air- or moisture-sensitive reagents wereperformed under a nitrogen atmosphere using dried solvents andglassware.

Analytical Conditions

Analytical Method # Description Analytical method 1 Solvents:Acetonitrile (far UV grade) with 0.1% (v/v) formic acid. Water (highpurity via PureLab Option unit) with 0.1% formic acid Column: PhenomenexLuna 5 μm C18 (2), 100 × 4.6 mm (Plus guard cartridge) Flow Rate: 2mL/min gradient: A: Water/formic acid B: MeCN/formic acid Time A % B %0.00 95 5 3.50 5 95 5.50 5 95 5.60 95 5 6.50 95 5 Typical Injections 2-7μL (concentration ~0.2-1.0 mg/mL) Analytical method 2 Solvents: -Acetonitrile (Far UV grade) Water (High purity via PureLab Option unit)with 10 mM ammonium bicarbonate (ammonium hydrogen carbonate) Column: -Waters Xterra MS 5μ C18, 100 × 4.6 mm (Plus guard cartridge) FlowRate: - 2 mL/min Gradient: - A: Water/Bicarb B: MeCN Time A % B % 0.0095 5 0.50 95 5 4.00 5 95 5.50 5 95 5.60 95 5 6.50 95 5 TypicalInjections 2-7 μL (concentration ~0.2-1 mg/mL) Analytical method 3Solvents: Acetonitrile (Far UV grade) with 0.1% (V/V) formic acid Water(High purity via PureLab Ultra unit) with 0.1% formic acid Column:Hichrom ACE 3 C18-AR mixed mode column 100 × 4.6 mm Flow Rate: 1 mL/mingradient: A: Water/formic B: MeCN/formic Time A % B % 0.00 98 2 3.00 982 12.00 0 100 15.4 0 100 15.5 98 2 17 98 2 Typical Injections 0.2-10 μL(concentration ~0.2-1.0 mg/mL) Analytical method 4 Solvents: -Acetonitrile (Far UV grade) with 0.1% (V/V) formic acid Water (Highpurity via PureLab Ultra unit) with 0.1% formic acid Column: Supelco,Ascentis ® Express C18 or Hichrom Halo C18, 2.7 μm C18, 150 × 4.6 mm.Both latest technology fused core columns Flow Rate: 1 ml/min Gradient:A: Water/formic B: MeCN/formic Time A % B % 0.00 96 4 3.00 96 4 9.00 0100 13.6 0 100 13.7 96 4 15 96 4 Typical Injections 0.2-10 μL(concentration ~0.2-1.0 mg/mL)General Synthetic Methods

Method a (Amide Coupling)

To a solution of carboxylic acid (1.50 mmol) in DCM (10 mL) at r.t. wereadded EDC (351 mg, 1.83 mmol) and HOPO (203 mg, 1.83 mmol). The mixturewas stirred for 30 min to give a complete solution then amine (free baseor hydrochloride salt) (1.65 mmol) and DIPEA (1.3 mL, 7.5 mmol) wereadded and the mixture stirred at r.t. for 18 h. The mixture was washedwith water, passed through a phase separation cartridge andconcentrated.

Method B (Amide Reduction)

To a solution of amide (1.30 mmol) in diethyl ether (50 mL) at −15 OCunder nitrogen was added dropwise lithium aluminum hydride (1.30 mL, 2.6mmol, 2.0 M in THF) over 20 min. The reaction was maintained at −10° C.for 1.5 h then quenched with water (1.5 mL), 2 N NaOH (1 mL) and furtherwater (2 mL). The reaction was warmed to r.t. and stirred for 30 min,then MgSO₄ was added and the reaction was filtered through Celite,washing well with EtOAc. The filtrate was concentrated and the residuepurified by passage through an SCX cartridge (5 g), eluting with 0-10% 7M methanolic ammonia in DCM.

Method C (Boc Removal)

To a solution of Boc-protected amine (0.76 mmol) in DCM (5 mL) at 0° C.under nitrogen was added dropwise 4 N HCl in dioxane (0.8 mL, 3.0 mmol).The solution was warmed to r.t. and stirred for 1 h, then concentrated.

Step 1: (R)-2-((tert-butoxycarbonyl)amino)propyl methanesulfonate(Intermediate 1)

A solution of methanesulfonyl chloride (7.29 mL, 94.2 mmol) in dry DCM(140 mL) was added dropwise, over a period of 1.5 h, to a solution of(R)-tert-butyl (1-hydroxypropan-2-yl)carbamate (15 g, 85.6 mmol) andtriethylamine (17.9 mL, 128.4 mmol) in DCM (300 mL) at 0° C. Thereaction was stirred at 0° C. for 30 min, then 1 h at r.t, beforewashing with water (300 mL), NaHCO₃ (300 mL) and brine (200 mL). Theorganics were passed through a phase separator, and concentrated underreduced pressure to give the title compound as a salmon pink solid (21.5g, 99%).

Method D

Amine (7.12 mmol) was added to intermediate 1 (900 mg, 3.56 mmol) andcesium carbonate (3.48 g, 10.7 mmol) in DMF (8 mL) and the mixturestirred at 65° C. for 24 h. The reaction mixture was cooled to r.t. andfiltered through celite, washing with MeOH. The filtrate wasconcentrated and the residue purified by passage through an SCXcartridge (5 g), eluting with 0-10% 7 M methanolic ammonia in DCM.

(R)-1-((S)-2-Methylpyrrolidin-1-yl)propan-2-amine dihydrochloride(Intermediate 2)

Step 1: tert-Butyl((R)-1-((S)-2-methylpyrrolidin-1-yl)-1-oxopropan-2-yl)carbamate

Following method A from (R)-2-((tert-butoxycarbonyl)amino)propanoic acid(1.14 g, 6.04 mmol) and(S)-2-methylpyrrolidine (565 mg, 6.65 mmol) gavethe title compound as a yellow oil which was progressed to the next stepas crude.

Step 2: tert-Butyl((R)-1-((S)-2-methylpyrrolidin-1-yl)propan-2-yl)carbamate

Following method B from tert-butyl((R)-1-((S)-2-methylpyrrolidin-1-yl)-1-oxopropan-2-yl)carbamate (1.55 g,6.04 mmol) gave the title compound as a colorless oil (1.43 g, 97%).

Step 3: (R)-1-((S)-2-Methylpyrrolidin-1-yl)propan-2-aminedihydrochloride (Intermediate 2)

Following method C from tert-butyl((R)-1-((S)-2-methylpyrrolidin-1-yl)propan-2-yl)carbamate (1.43 g, 5.88mmol) gave the title compound as a colorless oil (850 mg, 81%).

Example 1:(2S)-2-Methyl-1-((R)-2-(3-methyl-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamido)propyl)pyrrolidin-1-iumformate

Step 1: (Z)-4-(N-Hydroxycarbamimidoyl)-3-methylbenzoic acid

To a stirred solution of 4-cyano-3-methylbenzoic acid (1.00 g, 6.21mmol) in EtOH (30 mL) was added NH₂OH.HCl (604 mg, 8.70 mmol) and KOH(1.04 g, 18.6 mmol). The mixture was stirred at r.t. for 17 h, thenneutralized with 1 M HCl_((aq)) and extracted into EtOAc (3×25 mL). Thecombined organics were dried (MgSO₄) and concentrated to give the titlecompound as a yellow solid (1.3 g, 108%).

Step 2: 3-Methyl-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoicacid

To a stirred solution of (Z)-4-(N′-hydroxycarbamimidoyl)-3-methylbenzoicacid (1.2 g, 6.21 mmol) in THF (30 mL) was added TFAA (1.29 mL, 9.32mmol). The mixture was stirred for 3 h, after which time an additional 5eqs of TFAA were added. After a further 2 h stirring, LCMS indicated thereaction was complete. The mixture was poured onto ice-water andacidified to pH 4 with 1 M HCl_((aq)), before extracting into EtOAc(3×30 mL). The combined organics were dried (MgSO₄) and concentrated.Purification by flash column chromatography (gradient elution i-hex [+3%AcOH] to 4:1 i-hex:EtOAc [+3% AcOH]) gave the title compound as anoff-white solid. LCMS (ES+) consistent with target (M+H)⁺.

Step 3:(2S)-2-Methyl-1-((R)-2-(3-methyl-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamido)propyl)pyrrolidin-1-iumformate

Following method A from3-methyl-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (490mg, 1.80 mmol) and intermediate 2(386 mg, 2.70 mmol). Purification byreverse phase chromatography gave the title compound as an orange gum(65 mg, 9%). LCMS (ES+) 397 (M+H)⁺, RT 2.85 min (Analytical method 1).¹H NMR δ (ppm) (400 MHz, DMSO-d₆) 8.40 (1H, d, J=8.3 Hz), 8.17 (1H, s),8.04 (1H, d, J=8.2 Hz), 7.92 (1H, s), 7.88-7.85 (1H, m), 4.19-4.10 (1H,m), 3.16-3.10 (1H, m), 2.78-2.67 (1H, m), 2.65 (3H, s), 2.42-2.33 (1H,m), 2.29-2.15 (2H, m), 1.93-1.83 (1H, m), 1.72-1.62 (2H, m), 1.36-1.25(1H, m), 1.19 (3H, d, J=6.7 Hz), 1.05 (3H, d, J=6.0 Hz). NH⁺ notobserved.

Example 2:N-(2-(3-Azabicyclo[3.2.1]octan-3-yl)ethyl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Step 1:N-(2-Hydroxyethyl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Following method A from4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (1.5 g, 5.8mmol) and 2-aminoethanol (420 μL, 6.96 mmol) gave the title compound asa white solid (938 mg, 54%). LCMS (ES+) consistent with target (M+H)⁺.

Step 2:N-(2-Oxoethyl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

To a suspension ofN-(2-hydroxyethyl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide(938 mg, 3.11 mmol) in DCM (50 mL) was added Dess-Martin periodinane(1.98 g, 4.67 mmol). The reaction mixture was stirred at r.t. for 66 hand then quenched with MeOH (15 mL). The mixture was stirred for 15 minand then concentrated onto silica. Purification by column chromatography(i-hex to EtOAc/i-hex (1:1)) gave the title compound (450 mg, 48%).

Step3:N-(2-((1R,5S)-3-Azabicyclo[3.2.1]octan-3-yl)ethyl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

To a stirred solution ofN-(2-oxoethyl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide (87mg, 0.4 mmol) in DCM:AcOH (10:1, 4 mL) was added3-azabicyclo[3.2.1]octane (70 mg, 0.48 mmol) and PS-triethylammoniumcyanoborohydride (200 mg, 0.8 mmol). The reaction was shaken for 4 hbefore filtering and concentrating under reduced pressure. Purificationby preparative-HPLC gave the title compound. LCMS (ES+) 395 (M+H)⁺, RT2.75 min (Analytical Method 1); ¹H NMR δ (ppm) (400 MHz, DMSO-d₆)8.55-8.50 (1H, m,), 8.17 (2H, d, J=8.5 Hz), 8.04 (2H, d, J=8.5 Hz),3.40-3.35 (2H, m), 2.72-2.66 (2H, m), 2.46 (2H, dd, J=6.8, 6.8 Hz),2.09-2.03 (4H, m), 1.60-1.51 (2H, m), 1.50-1.38 (3H, m), 1.31 (1H, d,J=10.7 Hz).

Example 3:(R)—N-(1-(Isoindolin-2-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Step 1: (R)-tert-Butyl (1-(isoindolin-2-yl)-1-oxopropan-2-yl)carbamate

Following method A from (R)-2-((tert-butoxycarbonyl)amino)propanoic acid(3.78 mmol) and isoindoline (4.16 mmol). Purification by columnchromatography (gradient elution, 0-100% EtOAc in i-hex) gave the titlecompound as a pale yellow gum (1.01 g, 87%, 60% pure) which was usedwithout further purification.

Step 2: (R)-tert-Butyl (1-(isoindolin-2-yl)propan-2-yl)carbamate

Following method B from (R)-tert-butyl(1-(isoindolin-2-yl)-1-oxopropan-2-yl)carbamate (3.78 mmol). The titlecompound was obtained as a yellow gum (0.84 g, 80%).

Step 3: (R)-1-(Isoindolin-2-yl)propan-2-amine dihydrochloride

Following method C from (R)-tert-butyl(1-(isoindolin-2-yl)propan-2-yl)carbamate (3.04 mmol). The titlecompound was obtained as a dark green foam (655 mg, 86%).

Step 4:(R)—N-(1-(Isoindolin-2-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Following method A from (R)-1-(isoindolin-2-yl)propan-2-aminedihydrochloride (2.53 mmol) and4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (2.78 mmol).Purification by preparative HPLC gave the title compound as a pale brownsolid (60 mg, 6%). LCMS (ES+) 417 (M+H)⁺, RT 9.72 min (Analytical Method3); ¹H NMR δ (ppm) (400 MHz, DMSO-d₆) 8.50 (1H, d, J=8.2 Hz), 8.17 (2H,d, J=8.5 Hz), 8.08 (2H, d, J=8.5 Hz), 7.25-7.16 (4H, m), 4.33-4.24 (1H,m), 3.98-3.88 (4H, m), 2.89 (1H, dd, J=7.8, 11.9 Hz), 2.73 (1H, dd,J=6.4, 11.9 Hz), 1.23 (3H, d, J=6.7 Hz).

Example 4:N-((2R)-1-(3-Azabicyclo[3.1.0]hexan-3-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Step 1: 3-Benzyl-3-azabicyclo[3.1.0]hexane

3-Benzyl-3-azabicyclo[3.1.0]hexane-2,4-dione (1.95 g, 9.69 mmol, CAS No.73799-63-0) was suspended in THF (11 mL) and cooled to 0° C. under anitrogen atmosphere. LiAlH₄ (2 M in THF, 19.5 mL, 39.0 mmol) was addedover several minutes with stirring. The reaction was heated to refluxand left to stir for 18 h. The reaction was then quenched with the slowaddition of water (3 mL), 2 N NaOH (4 mL) and more water (8 mL). MgSO₄was added and the mixture stirred for 30 min before being filteredthrough Celite and washed with EtOAc. The solvents were removed underreduced pressure to afford the title compound as an oily yellow solid(1.38 g, 82%). The compound was used in the next step without furtherpurification.

Step 2: 3-Azabicyclo[3.1.0]hexane hydrochloride

3-Benzyl-3-azabicyclo[3.1.0]hexane (1.1 g, 6.35 mmol) was dissolved inmethanol (40 mL) and 10% Pd/C (300 mg, 0.282 mmol) was added. Themixture was shaken under a hydrogen gas atmosphere (2 bar) for 18 h atr.t. The reaction mixture was then filtered through Celite, acidifiedwith 4 N HCl in dioxane (2.38 mL, 9.53 mmol) and concentrated to yieldthe title compound a brown solid (680 mg, 90%) which was used in thenext step without further purification.

Step 3: tert-Butyl((2R)-1-(3-azabicyclo[3.1.0]hexan-3-yl)-1-oxopropan-2-yl)carbamate

3-Azabicyclo[3.1.0]hexane hydrochloride (670 mg, 5.6 mmol) was dissolvedin DMF (15 mL) and HATU (2.55 g, 6.70 mmol) and Boc-D-Alanine (1.06 g,5.60 mmol) added and the mixture stirred at r.t. for 10 minutes. DIPEA(2.92 mL, 16.8 mmol) was then added and the mixture left to stir at r.t.for 18 h before being diluted with EtOAc (20 mL), washed with 1 M HCl(15 mL), saturated NaHCO₃ (15 mL) and brine (15 mL). The organic layerwas dried over MgSO₄, filtered and the filtrate concentrated to yield ayellow oil Purification by column chromatography (gradient elution,i-hex to 60% EtOAc in i-hex) to yield the title compound as a clear oil(380 mg, 27%).

Step 4: tert-Butyl((2R)-1-(3-azabicyclo[3.1.0]hexan-3-yl)propan-2-yl)carbamate

Following method B from tert-butyl((2R)-1-(3-azabicyclo[3.1.0]hexan-3-yl)-1-oxopropan-2-yl)carbamate (380mg, 1.50 mmol) afforded the title compound as a yellow oil (290 mg,80%). The compound was used in the next step without furtherpurification.

Step 5: (2R)-1-(3-Azabicyclo[3.1.0]hexan-3-yl)propan-2-amine

Following method C from tert-butyl((2R)-1-(3-azabicyclo[3.1.0]hexan-3-yl)propan-2-yl)carbamate (290 mg,1.20 mmol). The reaction mixture was then concentrated to yield a brownoil which was purified by SCX cartridge, eluting with MeOH/DCM (1:1) andthen 0.5 N NH₃ in MeOH to yield the title compound as a yellow oil (110mg, 65%) which was used in the next step without further purification.

Step 6:N-((2R)-1-(3-Azabicyclo[3.1.0]hexan-3-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Following method A from(2R)-1-(3-azabicyclo[3.1.0]hexan-3-yl)propan-2-amine (83 mg, 0.59 mmol)and 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (153 mg,0.59 mmol). Purification by preparative-HPLC gave the title compound asan off white solid. LCMS (ES+) 381 (M+H)⁺, RT 2.61 min (AnalyticalMethod 1); ¹H NMR δ (ppm) (400 MHz, CDCl₃) 8.19 (2H, td, J=1.8, 8.4 Hz),7.92 (2H, td, J=1.8, 8.5 Hz), 6.97 (1H, br s), 4.08 (1H, br s), 3.10(1H, d, J=8.8 Hz), 3.03 (1H, d, J=8.8 Hz), 2.74 (1H, t, J=10.3 Hz), 2.58(1H, d, J=8.4 Hz), 2.52 (1H, dd, J=5.2, 12.3 Hz), 2.41 (1H, d, J=8.1Hz), 1.42-1.39 (2H, m), 1.31 (3H, d, J=6.4 Hz), 0.65 (1H, dd, J=3.9, 8.0Hz), 0.40 (1H, dt, J=4.3, 7.7 Hz).

Example 5:(S)—N-(1-Cyclopropyl-2-(pyrrolidin-1-yl)ethyl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Step 1: (S)-tert-Butyl(1-cyclopropyl-2-oxo-2-(pyrrolidin-1-yl)ethyl)carbamate

Following method A from(S)-2-((tert-butoxycarbonyl)amino)-2-cyclopropylacetic acid (1.0 g, 4.65mmol) and pyrrolidine (0.58 mL, 6.98 mmol) gave the title compound as acolorless oil which was progressed to the next step without furtherpurification.

Step 2: (S)-tert-Butyl (1-cyclopropyl-2-(pyrrolidin-1-yl)ethyl)carbamate

Following method B from (S)-tert-butyl(1-cyclopropyl-2-oxo-2-(pyrrolidin-1-yl)ethyl)carbamate (1.53 g, 5.7mmol) gave the title compound which was used without furtherpurification.

Step 3: (S)-1-Cyclopropyl-2-(pyrrolidin-1-yl)ethanamine

Following method C from(S)-tert-butyl(1-cyclopropyl-2-(pyrrolidin-1-yl)ethyl)carbamate (920 mg, 3.62 mmol).The free amine was isolated by dissolving the residue obtained afteraqueous work-up in DCM/MeOH (1:1, 5 mL) and passing it through an SCXcartridge (elution with DCM:MeOH:7M NH₃ in MeOH (1:1:0.05). The titlecompound was obtained as an orange oil (390 mg, 70%).

Step 4:(S)—N-(1-Cyclopropyl-2-(pyrrolidin-1-yl)ethyl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Following method A from (S)-1-cyclopropyl-2-(pyrrolidin-1-yl)ethanamine(134 mg, 0.87 mmol) and4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (150 mg, 0.58mmol). LCMS (ES+) 395 (M+H)⁺, RT 2.67 min (Analytical method 1). ¹H NMRδ (ppm) (400 MHz, DMSO-d₆) 8.42 (1H, d, J=8.7 Hz), 8.15 (2H, d, J=8.4Hz), 8.05 (2H, d, J=8.5 Hz), 3.71-3.63 (1H, m), 2.80 (1H, dd, J=9.1,12.1 Hz), 2.52-2.49 (3H, m), 2.48-2.40 (2H, m), 1.62 (4H, s), 1.02-0.93(1H, m), 0.50-0.43 (1H, m), 0.36-0.29 (2H, m), 0.24-0.17 (1H, m).

Example 6:(R)—N-(1-Cyclopropyl-2-(pyrrolidin-1-yl)ethyl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Step 1: (R)-tert-Butyl(1-cyclopropyl-2-oxo-2-(pyrrolidin-1-yl)ethyl)carbamate

Following method A from(R)-2-((tert-butoxycarbonyl)amino)-2-cyclopropylacetic acid (1.64 g,7.62 mmol) and pyrrolidine (0.95 mL, 11.4 mmol) gave the title compoundas a colorless oil which was progressed to the next step without furtherpurification.

Step 2: (R)-tert-Butyl (1-cyclopropyl-2-(pyrrolidin-1-yl)ethyl)carbamate

Following method B from (R)-tert-butyl(1-cyclopropyl-2-oxo-2-(pyrrolidin-1-yl)ethyl)carbamate (1.02 g, 3.81mmol) gave the title compound which was used without furtherpurification.

Step 3: (R)-1-Cyclopropyl-2-(pyrrolidin-1-yl)ethanamine

Following method C from (R)-tert-butyl(1-cyclopropyl-2-(pyrrolidin-1-yl)ethyl)carbamate (860 mg, 3.39 mmol).The free amine was isolated by dissolving the residue obtained afteraqueous work-up in DCM/MeOH (1:1, 5 mL) and passing it through an SCXcartridge (elution with DCM:MeOH:7M NH₃ in MeOH (1:1:0.05). The titlecompound was obtained as an orange oil (190 mg, 36%).

Step 4:(R)—N-(1-Cyclopropyl-2-(pyrrolidin-1-yl)ethyl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Following method A from (R)-1-cyclopropyl-2-(pyrrolidin-1-yl)ethanamine(190 mg, 1.23 mmol) and4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (212 mg, 0.82mmol). LCMS (ES+) 395 (M+H)⁺, RT 2.69 min (Analytical method 1). ¹H NMRδ (ppm) (400 MHz, DMSO-d₆) 8.42 (1H, d, J=8.7 Hz), 8.15 (2H, d, J=8.4Hz), 8.05 (2H, d, J=8.5 Hz), 3.71-3.63 (1H, m), 2.80 (1H, dd, J=9.1,12.1 Hz), 2.52-2.49 (3H, m), 2.48-2.40 (2H, m), 1.62 (4H, s), 1.02-0.93(1H, m), 0.50-0.43 (1H, m), 0.36-0.29 (2H, m), 0.24-0.17 (1H, m).

Example 7:(R)—N-(1-(3,4-Dihydroisoquinolin-2(1H)-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Step 1: (R)-tert-Butyl(1-(3,4-dihydroisoquinolin-2(1H)-yl)-1-oxopropan-2-yl)carbamate

Following method A from (R)-2-((tert-butoxycarbonyl)amino)propanoic acid(720 mg, 3.78 mmol) and 1,2,3,4-tetrahydroisoquinoline (0.52 mL, 4.016mmol). Purification by column chromatography (gradient elution, 0 to100% EtOAc in i-hex) gave the title compound as a pale yellow gum (1.01g, 87%).

Step 2: (R)-tert-Butyl(1-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-yl)carbamate

Following method B from(1-(3,4-dihydroisoquinolin-2(1H)-yl)-1-oxopropan-2-yl)carbamate (1.01 g,3.31 mmol) gave the title compound as a pale yellow gum (0.86 mg, 89%).

Step 3: (R)-1-(3,4-Dihydroisoquinolin-2(1H)-yl)propan-2-aminedihydrochloride

Following method C from (R)-tert-butyl(1-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-yl)carbamate (860 mg, 2.96mmol) gave the title compound as a pale orange glass (788 mg, >99%).This was used in the next step without further purification.

Step 4:(R)—N-(1-(3,4-Dihydroisoquinolin-2(1H)-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Following method A from4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (636 mg, 2.46mmol) and (R)-1-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-aminedihydrochloride (778 mg, 2.96 mmol) and DIPEA (2.2 mL, 12.3 mmol) wereadded and the mixture stirred for 18 h. The mixture was washed withwater, passed through a phase separation cartridge and concentrated toyield the crude product. This residue was purified by preparative-HPLCand then further purified by chiral SFC to give the title compound as awhite solid. LCMS (ES+) 431 (M+H)⁺, RT 2.80 min (Analytical Method 1);¹H NMR δ (ppm) (400 MHz, DMSO-d₆) 8.47 (1H, d, J=8.2 Hz), 8.16 (2H, d,J=8.5 Hz), 8.06 (2H, d, J=8.7 Hz), 7.12-7.04 (4H, m), 4.40-4.32 (1H, m),3.63 (2H, s), 2.82-2.62 (6H, m), 1.22 (3H, d, J=6.5 Hz).

Example 8:(R)—N-(1-(3-Phenylazetidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Step 1: (R)-tert-Butyl(1-oxo-1-(3-phenylazetidin-1-yl)propan-2-yl)carbamate

Following method A from (R)-2-((tert-butoxycarbonyl)amino)propanoic acid(649 mg, 3.41 mmol) 3-phenylazetidine (0.5 g, 3.75 mmol). Purificationby column chromatography (gradient elution, 0 to 100% EtOAc/i-hex) gavethe title compound as a colorless gum (0.87 g, 83%).

Step 2: (R)-tert-Butyl (1-(3-phenylazetidin-1-yl)propan-2-yl)carbamate

Following method B from (R)-tert-butyl(1-oxo-1-(3-phenylazetidin-1-yl)propan-2-yl)carbamate (0.87 g, 2.86mmol). Purification by column chromatography (gradient elution, 0 to100% EtOAc/i-hex) gave the title compound as a colorless oil (370 mg,44%).

Step 3: (R)-1-(3-Phenylazetidin-1-yl)propan-2-amine dihydrochloride

Following method C from (R)-tert-butyl(1-(3-phenylazetidin-1-yl)propan-2-yl)carbamate (370 mg, 1.27 mmol gavethe title compound as an off white solid (337 mg, >99%). The materialwas progressed to the next step without further purification.

Step 4:(R)—N-(1-(3-Phenylazetidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Following method A from4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (252 mg, 0.98mmol) and (R)-1-(3-phenylazetidin-1-yl)propan-2-amine dihydrochloride(348 mg, 1.07 mmol). After work up the crude material was recrystallizedfrom diisopropyl ether three times to give the title compound as a whitesolid (71 mg, 16%). LCMS (ES+) 431 (M+H)⁺, RT 2.83 min (AnalyticalMethod 1); ¹H NMR δ (ppm) (400 MHz, DMSO-d₆) 8.42 (1H, d, J=8.0 Hz),8.17 (2H, d, J=8.4 Hz), 8.07 (2H, d, J=8.5 Hz), 7.35-7.28 (4H, m),7.23-7.18 (1H, m), 4.11-4.00 (1H, m), 3.75-3.57 (3H, m), 3.20-3.07 (2H,m), 2.68-2.57 (1H, m), 1.18 (3H, d, J=6.8 Hz). One proton obscured byDMSO peak.

Example 9 and Example 10: D1:N—((R)-1-((abs)-3-(Difluoromethoxy)piperidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;and D2:N—((R)-1-((abs)-3-(difluoromethoxy)piperidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Step 1: tert-Butyl((2R)-1-(3-(difluoromethoxy)piperidin-1-yl)-1-oxopropan-2-yl)carbamate

Following Method A from (R)-2-((tert-butoxycarbonyl)amino)propanoic acid(284 mg, 1.50 mmol) and (±)-3-(difluoromethoxy)piperidine (250 mg, 1.65mmol). The title compound was obtained as a yellow gum (424 mg, 87%) andused in the next step without further purification.

Step 2: tert-Butyl((2R)-1-(3-(difluoromethoxy)piperidin-1-yl)propan-2-yl)carbamate

Following method B from tert-butyl((2R)-1-(3-(difluoromethoxy)piperidin-1-yl)-1-oxopropan-2-yl)carbamate(420 mg, 1.30 mmol). The title compound was obtained as a pale yellowgum (234 mg, 58%).

Step 3: (2R)-1-(3-(Difluoromethoxy)piperidin-1-yl)propan-2-aminedihydrochloride

Following method C from tert-butyl((2R)-1-(3-(difluoromethoxy)piperidin-1-yl)propan-2-yl)carbamate (234mg, 0.76 mmol). The title compound was obtained as a pale yellow foam(210 mg, 98%).

Step 4: D1:N—((R)-1-((abs)-3-(Difluoromethoxy)piperidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;and D2:N—((R)-1-((abs)-3-(difluoromethoxy)piperidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Following method A from4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (178 mg, 0.69mmol) and (2R)-1-(3-(difluoromethoxy)piperidin-1-yl)propan-2-aminedihydrochloride (210 mg, 0.76 mmol). The crude material was purified bypreparative HPLC and chiral preparative SFC (LUX Cellulose-4, 5/95IPA/CO₂, 5.0 ml/min, 120 bar, 40° C. to give the title compounds aswhite solids. D1:N—((R)-1-((abs)-3-(difluoromethoxy)piperidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide(50 mg, 16%): LCMS (ES+) 449 (M+H)⁺, RT 2.74 min (Analytical Method 1);SFC RT 4.48 min; ¹H NMR δ (ppm) (400 MHz, DMSO-d₆) 8.40 (1H, d, J=8.3Hz), 8.17 (2H, d, J=8.5 Hz), 8.05 (2H, d, J=8.5 Hz), 6.72 (1H, t, J=76.3Hz), 4.27-4.16 (1H, m), 4.10-4.01 (1H, m), 2.91 (1H, dd, J=3.0, 10.4Hz), 2.68-2.60 (1H, m), 2.35 (1H, dd, J=7.0, 12.4 Hz), 2.19-2.05 (2H,m), 1.91-1.82 (1H, m), 1.71-1.63 (1H, m), 1.47-1.27 (2H, m), 1.16 (3H,d, J=6.5 Hz). 1 proton obscured by DMSO peak. D2:N—((R)-1-((abs)-3-(difluoromethoxy)piperidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide(53 g, 17%): LCMS (ES+)449 (M+H)⁺, RT 2.72 min (Analytical Method 1);SFC RT 7.49 min; ¹H NMR δ (ppm) (400 MHz, DMSO-d₆) 8.40 (1H, d, J=8.2Hz), 8.17 (2H, d, J=8.5 Hz), 8.05 (2H, d, J=8.5 Hz), 6.73 (1H, t, J=76.4Hz), 4.27-4.16 (1H, m), 4.11-4.02 (1H, m), 2.92 (1H, dd, J=3.3, 10.6Hz), 2.72-2.67 (1H, m), 2.35 (1H, dd, J=6.7, 12.5 Hz), 2.15-2.02 (2H,m), 1.92-1.83 (1H, m), 1.70-1.62 (1H, m), 1.46-1.25 (2H, m), 1.16 (3H,d, J=6.7 Hz). 1 proton obscured by DMSO peak.

Example 11:(R)—N-(1-(5-Azaspiro[2.5]octan-5-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Step 1: (R)-tert-Butyl(1-oxo-1-(5-azaspiro[2.5]octan-5-yl)propan-2-yl)carbamate

To a stirred solution of (R)-2-((tert-butoxycarbonyl)amino)propanoicacid (648 mg, 3.43 mmol) in DMF (10 mL) was added HBTU (1.43 g, 3.73mmol), 5-azaspiro[2.5]octane hydrochloride (500 mg, 3.43 mmol) and DIPEA(3.16 mL, 17.1 mmol). The mixture was stirred at r.t. overnight. Themixture was washed with water, passed through a phase separationcartridge and concentrated to give the title compound (1.01 g) which wasused in the next step without further purification.

Step 2: (R)-tert-Butyl(1-(5-azaspiro[2.5]octan-5-yl)propan-2-yl)carbamate

Following method B from (R)-tert-butyl(1-oxo-1-(5-azaspiro[2.5]octan-5-yl)propan-2-yl)carbamate (1.01 g, 3.58mmol) gave the title compound which was used in the next step withoutfurther purification.

Step 3: (R)-1-(5-Azaspiro[2.5]octan-5-yl)propan-2-amine dihydrochloride

Following method C from (R)-tert-butyl(1-(5-azaspiro[2.5]octan-5-yl)propan-2-yl)carbamate (866 mg, 2.29 mmol)gave the title compound which was used in the next step without furtherpurification.

Step 4:(R)—N-(1-(5-Azaspiro[2.5]octan-5-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Using the same method as in step 1 from4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (590 mg, 2.29mmol) and (R)-1-(5-azaspiro[2.5]octan-5-yl)propan-2-aminedihydrochloride (770 mg, 2.29 mmol). The title compound was obtained asa white solid. LCMS (ES+) 409 (M+H)⁺, RT 3.58 min (Analytical Method 2);¹H NMR δ (ppm) (400 MHz, DMSO-d₆) 8.38 (1H, d, J=8.2 Hz), 8.19 (2H, d,J=8.5 Hz), 8.07 (2H, d, J=8.5 Hz), 4.24-4.16 (1H, m), 2.49-2.41 (2H, m),2.29 (1H, dd, J=7.2, 12.2 Hz), 2.18 (2H, dd, J=11.8, 17.5 Hz), 1.63-1.57(2H, m), 1.32-1.22 (3H, m), 1.17 (3H, d, J=6.5 Hz), 0.31-0.22 (4H, m).

Example 12:(R)—N-(1-(6-Azaspiro[2.5]octan-6-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Step 1: (R)-tert-Butyl(1-oxo-1-(6-azaspiro[2.5]octan-6-yl)propan-2-yl)carbamate

A flask was charged with (R)-2-((tert-butoxycarbonyl)amino)propanoicacid (250 mg, 1.32 mmol), 6-azaspiro[2.5]octane (150 mg, 1.32 mmol),HATU (602 mg, 1.60 mmol) and anhydrous DMF (5 mL). The reaction mixturewas stirred until a pale yellow solution formed and then DIPEA (350 μL,2.0 mmol) was added and the mixture stirred overnight. The reactionmixture was diluted with EtOAc (25 mL), washed with 1 M HCl (15 mL),sat. NaHCO₃ (15 mL) then brine (15 mL). The organics were dried (MgSO₄),filtered and concentrated. Purification by column chromatography(gradient elution, 0 to 66% EtOAc in i-hex) gave the title compound as acolorless oil (339 mg, 91%) which solidifies on standing.

Step 2: (R)-tert-Butyl(1-(6-azaspiro[2.5]octan-6-yl)propan-2-yl)carbamate

Following method B (using THF instead of Et₂O) from (R)-tert-butyl(1-oxo-1-(6-azaspiro[2.5]octan-6-yl)propan-2-yl)carbamate (320 mg, 1.13mmol) gave the title compound as a pale yellow oil (260 mg, 86%).

Step 3: (R)-1-(6-Azaspiro[2.5]octan-6-yl)propan-2-amine

To a solution of (R)-tert-butyl(1-(6-azaspiro[2.5]octan-6-yl)propan-2-yl)carbamate (220 mg, 0.82 mmol)in anhydrous DCM (10 mL) was added TFA (1.5 mL). The reaction mixturewas stirred at r.t. for 48 h. The mixture was concentrated under reducedpressure and passed through an SCX cartridge (elution with MeOH:7 M NH₃in MeOH (4:1)). The basic fractions were concentrated to give the titlecompound as a brown oil (120 mg, 85%).

Step 4:(R)—N-(1-(6-Azaspiro[2.5]octan-6-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Following the same method as for step 1 from4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (124 mg, 0.48mmol) and (R)-1-(6-azaspiro[2.5]octan-6-yl)propan-2-amine (80 mg, 0.48mmol). Purification by preparative HPLC gave the title compound as ancolorless solid. LCMS (ES+) 409 (M+H)⁺, RT 2.77 min (Analytical Method1); ¹H NMR δ (ppm) (400 MHz, DMSO-d₆) 8.16 (1H, d, J=8.1 Hz), 7.96-7.90(2H, m), 7.82 (2H, d, J=8.3 Hz), 4.00 (1H, t, J=7.2 Hz), 3.10 (4H, brs), 2.2.30-2.20 (5H, s), 2.14-2.08 (1H, m), 1.08 (4H, s), 0.93 (3H, t,J=6.6 Hz).

Example 13:(R)—N-(1-(Azepan-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Step 1: (R)-tert-Butyl (1-(azepan-1-yl)propan-2-yl)carbamate

Following method D from intermediate 1 (1.0 g, 3.95 mmol) and azepane(890 μL, 7.91 mmol) gave the title compound as a yellow oil (489 mg,48%).

Step 2: (R)-1-(Azepan-1-yl)propan-2-amine

To a solution of (R)-tert-butyl (1-(azepan-1-yl)propan-2-yl)carbamate(489 mg, 1.91 mmol) in anhydrous DCM (40 mL) was added TFA (0.8 mL). Thereaction mixture was stirred at r.t. for 2 h. The mixture wasconcentrated under reduced pressure and passed through an SCX cartridge(elution with MeOH:7 M NH₃ in MeOH (4:1)). The basic fractions wereconcentrated to give the title compound as a yellow oil (310 mg, >99%).

Step 3:(R)—N-(1-(Azepan-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Following method A from4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (427 mg, 1.65mmol) and (R)-1-(azepan-1-yl)propan-2-amine (310 mg, 1.91 mmol).Purification by preparative-HPLC and SFC gave the title compound as awhite solid (45 mg). LCMS (ES+) 397 (M+H)⁺, RT 2.70 min (AnalyticalMethod 1); ¹H NMR δ (ppm) (400 MHz, DMSO-d₆) 8.35 (1H, d, J=8.2 Hz),8.17 (2H, d, J=8.7 Hz), 8.05 (2H, d, J=8.5 Hz), 4.17-4.09 (1H, m),2.68-2.59 (5H, m), 2.45 (1H, dd, J=7.0, 12.4 Hz), 1.58-1.51 (8H, m),1.16 (3H, d, J=6.7 Hz).

Example 14:(R)—N-(1-(1-Azaspiro[3.3]heptan-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Step 1: (R)-tert-butyl(1-(1-Azaspiro[3.3]heptan-1-yl)propan-2-yl)carbamate

Following method D from intermediate 1 (326 mg, 1.29 mmol) and1-azaspiro[3.3]heptane (250 mg, 2.53 mmol) gave the title compound (250mg) which was used in the next step without further purification.

Step 2: 1-((R)-2-Aminopropyl)-1-azaspiro[3.3]heptan-1-ium2,2,2-trifluoroacetate

To a solution of (R)-tert-butyl(1-(1-azaspiro[3.3]heptan-1-yl)propan-2-yl)carbamate (0.25 mg, 1.00mmol) in anhydrous DCM (2 mL) was added TFA (1 mL). The reaction mixturewas stirred at r.t. for 3 h. The mixture was concentrated under reducedpressure to give the title compound (268 mg) which was used in the nextstep without further purification.

Step 3:(R)—N-(1-(1-Azaspiro[3.3]heptan-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

A flask was charged with (R)-2-((tert-butoxycarbonyl)amino)propanoicacid (260 mg, 1.00 mmol),1-((R)-2-aminopropyl)-1-azaspiro[3.3]heptan-1-ium 2,2,2-trifluoroacetate(268 mg, 1.00 mmol), HATU (690 mg, 1.5 mmol), DMAP (30 mg) and anhydrousDMF (5 mL). The reaction mixture was stirred until a pale yellowsolution formed and then DIPEA (1.5 mL, 72.0 mmol) was added and themixture stirred overnight. The reaction mixture was diluted with DCM,washed with water passed through a phase separator, filtered andconcentrated. Purification by preparative-HPLC and SFC gave the titlecompound as a white solid (12 mg). LCMS (ES+) 395 (M+H)⁺, RT 2.65 min(Analytical Method 1); ¹H NMR δ (ppm) (400 MHz, DMSO-d₆) 8.42 (1H, d,J=8.1 Hz), 8.21 (2H, d, J=8.6 Hz), 8.11 (2H, d, J=8.6 Hz), 4.11-4.02(1H, m), 3.15-3.08 (2H, m), 2.63 (1H, dd, J=6.4, 11.7 Hz), 2.49 (1H, dd,J=7.1, 11.9 Hz), 2.27-2.09 (4H, m), 1.90 (2H, m), 1.64-1.56 (2H, m),1.21 (3H, d, J=6.8 Hz).

Example 15:(R)—N-(1-(5-Azaspiro[2.4]heptan-5-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Step 1: (R)-tert-butyl(1-(5-Azaspiro[2.4]heptan-5-yl)propan-2-yl)carbamate

Following method D from intermediate 1 (237 mg, 0.90 mmol) and5-azaspiro[2.4]heptan-5-ium chloride (250 mg, 1.80 mmol) gave the titlecompound (100 mg) which was used in the next step without furtherpurification.

Step 2: 5-((R)-2-Aminopropyl)-5-azaspiro[2.4]heptan-5-ium2,2,2-trifluoroacetate

To a solution of (R)-tert-butyl(1-(1-azaspiro[3.3]heptan-1-yl)propan-2-yl)carbamate (0.25 mg, 1.00mmol) in anhydrous DCM (2 mL) was added TFA (1 mL). The reaction mixturewas stirred at r.t. for 3 h. The mixture was concentrated under reducedpressure to give the title compound (268 mg) which was used in the nextstep without further purification.

Step 3:(R)—N-(1-(1-Azaspiro[3.3]heptan-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

A flask was charged with (R)-2-((tert-butoxycarbonyl)amino)propanoicacid (260 mg, 1.00 mmol),(R)-1-(1-azaspiro[3.3]heptan-1-yl)propan-2-amine (268 mg, 1.00 mmol),HATU (690 mg, 1.5 mmol), DMAP (30 mg) and anhydrous DMF (5 mL). Thereaction mixture was stirred until a pale yellow solution formed andthen DIPEA (1.5 mL, 72.0 mmol) was added and the mixture stirredovernight. The reaction mixture was diluted with DCM, washed with waterpassed through a phase separator, filtered and concentrated.Purification by preparative-HPLC and SFC gave the title compound as awhite solid (12 mg). LCMS (ES+) 395 (M+H)⁺, RT 2.65 min (AnalyticalMethod 1); ¹H NMR δ (ppm) (400 MHz, DMSO-d₆) 8.48 (1H, d, J=8.1 Hz),8.22 (2H, d, J=8.6 Hz), 8.12 (2H, d, J=8.3 Hz), 4.24-4.16 (1H, m), 2.73(2H, dd, J=6.9, 6.9 Hz), 2.62 (2H, dd, J=7.3, 11.6 Hz), 2.52-2.48 (2H,m), 1.79-1.74 (2H, m), 1.23 (3H, d, J=6.6 Hz), 0.54 (4H, dd, J=7.6, 18.9Hz).

Example 16:N-((2R)-1-(3-Azabicyclo[3.2.1]octan-3-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Step 1: tert-Butyl((R)-1-((1R,5S)-3-azabicyclo[3.2.1]octan-3-yl)propan-2-yl)carbamate

Following method D from intermediate 1 (736 mg, 2.90 mmol) and(1R,5S)-3-azabicyclo[3.2.1]octane (730 mg, 4.94 mmol) gave the titlecompound (186 mg, 21%) which was used in the next step without furtherpurification.

Step 2: (R)-1-((1R,5S)-3-Azabicyclo[3.2.1]octan-3-yl)propan-2-amine

To a solution of tert-butyl((R)-1-((1R,5S)-3-azabicyclo[3.2.1]octan-3-yl)propan-2-yl)carbamate (186mg, 0.62 mmol) in anhydrous DCM (2.5 mL) was added TFA (0.5 mL). Thereaction mixture was stirred at r.t. for 3 h. The mixture wasconcentrated under reduced pressure and passed through an SCX cartridge(elution with MeOH:7 M NH₃ in MeOH (4:1)). The basic fractions wereconcentrated to give the title compound (80 mg) which was used in thenext step without further purification.

Step 3:N—((R)-1-((1R,5S)-3-Azabicyclo[3.2.1]octan-3-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Following method A from4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (122 mg, 0.47mmol) and (R)-1-(3-azabicyclo[3.2.1]octan-3-yl)propan-2-amine (80 mg,0.47 mmol).

Purification by preparative-HPLC and SFC gave the title compound as awhite solid (33 mg). LCMS (ES+) 409 (M+H)⁺, RT 2.81 min (AnalyticalMethod 1); ¹H NMR δ (ppm) (400 MHz, DMSO-d₆) 8.31 (1H, d, J=8.3 Hz),8.17 (2H, d, J=8.7 Hz), 8.05 (2H, d, J=8.5 Hz), 4.24-4.16 (1H, m),2.73-2.61 (2H, m), 2.41 (1H, dd, J=8.3, 12.2 Hz), 2.26 (1H, dd, J=6.7,12.2 Hz), 2.11-1.99 (4H, m), 1.55-1.36 (5H, m), 1.29 (1H, d, J=10.4 Hz),1.15 (3H, d, J=6.5 Hz).

Example 17:2-((R)-2-(4-(5-(Trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamido)propyl)octahydrocyclopenta[c]pyrrol-2-iumformate

Step 1: tert-Butyl((2R)-1-(hexahydrocyclopenta[c]pyrrol-2(1H)-yl)-1-oxopropan-2-yl)carbamate

Following method A from (R)-2-((tert-butoxycarbonyl)amino)propanoic acid(0.77 g, 4.09 mmol) and octahydrocyclopenta[c]pyrrole (0.50 g, 4.5 mmol)gave the title compound as a yellow gum (1.08 g, 93%). The compound wasimpure but used in the next step without further purification.

Step 2: tert-Butyl((2R)-1-(hexahydrocyclopenta[c]pyrrol-2(1H)-yl)-1-oxopropan-2-yl)carbamate

Following method B from tert-butyl((2R)-1-(hexahydrocyclopenta[c]pyrrol-2(1H)-yl)-1-oxopropan-2-yl)carbamate(1.08 g, 3.82 mmol) gave the title compound as a pale yellow gum (0.61g, 59%). The compound was impure but used in the next step withoutfurther purification.

Step 3: (2R)-1-(Hexahydrocyclopenta[c]pyrrol-2(1H)-yl)propan-2-aminehydrochloride

Following method C from tert-butyl((2R)-1-(hexahydrocyclopenta[c]pyrrol-2(1H)-yl)-1-oxopropan-2-yl)carbamate(0.61 g, 2.27 mmol) gave the title compound as a pale yellow foam (274mg, >100%). This was used in the next step without further purification.

Step 4:2-((R)-2-(4-(5-(Trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamido)propyl)octahydrocyclopenta[c]pyrrol-2-iumformate

Following method A from4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (0.48 g, 1.89mmol). Purification by column chromatography using gradient elution0-20% methanol in DCM gave a pale orange solid (700 mg). This waspurified further by preparative HPLC to give the title compound as apale orange glass (142 mg). LCMS (ES+) 409 (M+H)⁺, RT 2.74 min(Analytical Method 1); ¹H NMR δ (ppm) (400 MHz, DMSO-d₆) 8.40 (1H, d,J=8.2 Hz), 8.19-8.15 (3H, m), 8.05 (2H, d, J=8.5 Hz), 4.23-4.11 (1H, m),2.60 (2H, q, J=7.5 Hz), 2.49-2.43 (3H, m), 2.43-2.38 (1H, m), 2.25 (2H,ddd, J=3.9, 8.4, 12.0 Hz), 1.59-1.52 (3H, m), 1.41-1.29 (3H, m), 1.17(3H, d, J=6.7 Hz).

Example 18:(R)—N-(1-(3,4-dihydro-2,7-naphthyridin-2(1H)-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Step 1: (R)-tert-Butyl(1-(3,4-dihydro-2,6-naphthyridin-2(1H)-yl)-1-oxopropan-2-yl)carbamate

Following method A from (R)-2-((tert-butoxycarbonyl)amino)propanoic acid(1.28 g, 6.78 mmol) and 1,2,3,4-tetrahydro-2,6-naphthyridine (1.0 g,7.45 mmol). The title compound was obtained as a pale yellow gum (2.43g, >100%). This was used crude in the next step.

Step 2: (R)-tert-Butyl(1-(3,4-dihydro-2,6-naphthyridin-2(1H)-yl)propan-2-yl)carbamate

Following method B from (R)-tert-butyl(1-(3,4-dihydro-2,6-naphthyridin-2(1H)-yl)-1-oxopropan-2-yl)carbamate(2.43 g, 7.96 mmol). The mixture was purified by passing through an SCXcartridge (elution with MeOH:7 M NH₃ in MeOH (9:1)). This gave the titlecompound as a pale yellow gum (810 mg, 34%). The product was used in thenext step without further purification.

Step 3: (R)-1-(3,4-Dihydro-2,6-naphthyridin-2(1H)-yl)propan-2-aminetrihydrochloride

Following method C from (R)-tert-butyl(1-(3,4-dihydro-2,6-naphthyridin-2(1H)-yl)propan-2-yl)carbamate (810 mg,2.78 mmol) gave the title compound as a pale orange glass (0.98mg, >100%). This was used in the next step without further purification.

Step 4:(R)—N-(1-(3,4-Dihydro-2,7-naphthyridin-2(1H)-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Following method A from4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (650 mg, 2.5mmol) and (R)-1-(3,4-dihydro-2,6-naphthyridin-2(1H)-yl)propan-2-aminetrihydrochloride (980 mg, 2.78 mmol) and DIPEA (2.2 mL, 12.5 mmol) wereadded and the mixture stirred for 18 h. The mixture was washed withwater, passed through a phase separation cartridge and concentrated toyield the crude product. Purification by preparative-HPLC gave the titlecompound as an off white solid (162 mg). LCMS (ES+) 432 (M+H)⁺, RT 2.41min (Analytical Method 1); ¹H NMR δ (ppm) (400 MHz, DMSO-d₆) 8.48 (1H,d, J=8.2 Hz), 8.29 (1H, s), 8.26 (1H, d, J=5.0 Hz), 8.16 (3H, d, J=7.8Hz), 8.06 (2H, d, J=8.5 Hz), 7.11 (1H, d, J=5.1 Hz), 4.42-4.30 (1H, m),3.66 (2H, s), 2.81-2.64 (6H, m), 1.22 (3H, d, J=6.5 Hz).

Example19:N-((2R)-1-(3-Azabicyclo[3.2.0]heptan-3-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Step 1: 3-(4-Methoxybenzyl)-3-azabicyclo[3.2.0]heptane-2,4-dione

4-Methoxy benzylamine (1.04 mL, 7.93 mmol) was added dropwise at 0° C.to a solution of 3-oxabicyclo[3.2.0]heptane-2,4-dione (1.0 g, 7.93 mmol)in THF (3 mL). The semi solid mixture was warmed to 90° C. to remove theTHF and the solid slowly melted as the mixture was warmed to 180° C. for2 h. The solution was cooled to 60° C. and poured into iced DIPE toafford the title compound as a white solid (1.77 g, 91%), which wascollected by filtration.

Step 2: 3-(4-Methoxybenzyl)-3-azabicyclo[3.2.0]heptane

Following method B from3-(4-methoxybenzyl)-3-azabicyclo[3.2.0]heptane-2,4-dione (1.0 g, 4.1mmol). The reaction was refluxed for 2 h before work-up. This gave thetitle compound as a pale yellow gum (1.0 g). The product was used in thenext step without further purification.

Step 3: 3-Azabicyclo[3.2.0]heptane hydrochloride

3-(4-Methoxybenzyl)-3-azabicyclo[3.2.0]heptane (870 mg, 4.0 mmol) wasdissolved in MeOH (100 mL) and 10% Pd/C (870 mg) was added. The mixturewas shaken under a hydrogen gas atmosphere (2 bar) for 18 h at r.t. Thereaction mixture was then filtered through Celite, acidified with 4 NHCl in dioxane (5 mL) and concentrated to yield the title compound as anoff-white glass (590 mg) which was used in the next step without furtherpurification.

Step 4: tert-Butyl((2R)-1-(3-azabicyclo[3.2.0]heptan-3-yl)-1-oxopropan-2-yl)carbamate

Following method A from (R)-2-((tert-butoxycarbonyl)amino)propanoic acid(630 mg, 3.33 mmol) and 3-azabicyclo[3.2.0]heptane hydrochloride (590mg, 4.0 mmol). The title compound was obtained as a pale yellow gum (490mg, 55%). This was used crude in the next step.

Step 5: tert-Butyl((2R)-1-(3-azabicyclo[3.2.0]heptan-3-yl)propan-2-yl)carbamate

Following method B from tert-butyl((2R)-1-(3-azabicyclo[3.2.0]heptan-3-yl)-1-oxopropan-2-yl)carbamate(0.49 g, 1.80 mmol). The mixture was purified by passing through an SCXcartridge (elution with MeOH:7 M NH₃ in MeOH (9:1)). This gave the titlecompound as a pale yellow gum (229 mg, 49%).

Step 6: (2R)-1-(3-Azabicyclo[3.2.0]heptan-3-yl)propan-2-aminehydrochloride

Following method C from tert-butyl((2R)-1-(3-azabicyclo[3.2.0]heptan-3-yl)propan-2-yl)carbamate (490 mg,0.90 mmol) gave the title compound as a pale yellow solid (254mg, >100%). This was used in the next step without further purification.

Step 7:N-((2R)-1-(3-Azabicyclo[3.2.0]heptan-3-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Following method A from4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (211 mg, 0.82mmol) and (2R)-1-(3-azabicyclo[3.2.0]heptan-3-yl)propan-2-aminehydrochloride (254 mg, 0.90 mmol) and DIPEA (0.73 mL, 4.09 mmol) wereadded and the mixture stirred for 18 h. The mixture was washed withwater, passed through a phase separation cartridge and concentrated toyield the crude product. This residue was purified by preparative-HPLCand then further purified by chiral SFC to give the title compound as awhite solid (65 mg). LCMS (ES+) 395 (M+H)⁺, RT 2.71 min (AnalyticalMethod 1); ¹H NMR δ (ppm) (400 MHz, DMSO-d₆) 8.44 (1H, d, J=8.3 Hz),8.17 (2H, d, J=8.5 Hz), 8.06 (2H, d, J=8.7 Hz), 4.31-4.22 (1H, m), 2.83(2H, dd, J=9.0, 29.3 Hz), 2.75-2.66 (2H, m), 2.61 (1H, dd, J=7.7, 11.6Hz), 2.53-2.50 (1H, m, obscured by DMSO), 2.06-1.98 (4H, m), 1.61-1.56(2H, m), 1.25 (3H, d, J=6.5 Hz).

Example 20 and 21: D1:N—((R)-1-((abs-1,5-cis)-6-Azabicyclo[3.2.0]heptan-6-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;and D2:N—((R)-1-((abs-1,5-cis)-6-azabicyclo[3.2.0]heptan-6-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Step 1: 6-Azabicyclo[3.2.0]heptane hydrochloride

A solution of the azabicyclo[3.2.0]heptan-7-one (500 mg, 4.49 mmol) indiethyl ether (9 mL) was added dropwise over 0.5 h to a refluxingsolution of LiAlH₄ (5.6 mL, 11.2 mmol) in diethyl ether (36 mL). Thereaction mixture was refluxed for 24 h, cooled to 0° C., then water (3mL), 2 N NaOH solution (4 mL) and water (8 mL) were added sequentially.The reaction was warmed to r.t. and stirred for 30 min, then MgSO₄ wasadded and the reaction was filtered through Celite, washing well withEtOAc. The filtrate was acidified with 4 N HCl in dioxane (5 mL) andconcentrated to give the title compound as an opaque gum (270 mg, 45%).The crude material was taken on to the next step without furtherpurification.

Step 2: tert-Butyl((2R)-1-(6-azabicyclo[3.2.0]heptan-6-yl)-1-oxopropan-2-yl)carbamate

Following method A from (R)-2-((tert-butoxycarbonyl)amino)propanoic acid(1.28 mg, 6.78 mmol) and 6-azabicyclo[3.2.0]heptane hydrochloride (270mg, 2.78 mmol). The title compound was obtained as a pale yellow gum(0.61 mg, 89%). This was used crude in the next step.

Step 3: tert-Butyl((2R)-1-(bicyclo[3.2.0]heptan-6-yl)propan-2-yl)carbamate (twodiastereomers)

Following method B from tert-butyl((2R)-1-(6-azabicyclo[3.2.0]heptan-6-yl)-1-oxopropan-2-yl)carbamate(0.61 g, 2.27 mmol). The mixture was purified by passing through an SCXcartridge (elution with MeOH:7 M NH₃ in MeOH (9:1)). This gave a mixtureof two diastereomers of the title compound as a pale yellow gum (250 mg,43%).

Step 4: (2R)-1-(6-Azabicyclo[3.2.0]heptan-6-yl)propan-2-aminedihydrochloride (two diastereomers)

Following method C from tert-butyl((2R)-1-(bicyclo[3.2.0]heptan-6-yl)propan-2-yl)carbamate (250 mg, 0.97mmol) gave a mixture of two diastereomers of the title compound as apale yellow solid (257 mg). This was used in the next step withoutfurther purification.

Step 5: D1:N—((R)-1-((abs-1,5-cis)-6-Azabicyclo[3.2.0]heptan-6-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide;and D2:N—((R)-1-((abs-1,5-cis)-6-azabicyclo[3.2.0]heptan-6-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Following method A from4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (228 mg, 0.88mmol) and (2R)-1-(6-azabicyclo[3.2.0]heptan-6-yl)propan-2-aminedihydrochloride (257 mg, 0.98 mmol) and DIPEA (0.79 mL, 4.43 mmol) wereadded and the mixture stirred for 18 h. The mixture was washed withwater, passed through a phase separation cartridge and concentrated toyield the crude product. This residue was purified by preparative-HPLCand then further purified by chiral preparative SFC (LUX CELLUOSE-410/90 MeOH (0.1% DEA)/CO₂, 5.0 ml/min, 120 bar, 40° C.) to give thetitle compounds as white solids. D1:N—((R)-1-((abs-1,5-cis)-6-Azabicyclo[3.2.0]heptan-6-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide(8 mg). LCMS (ES+) 395 (M+H)⁺, RT 3.67 min (Analytical Method 2); ¹H NMRδ (ppm) (400 MHz, DMSO-d₆) 8.34 (1H, br s), 8.17 (2H, d, J=8.3 Hz), 8.05(2H, d, J=8.4 Hz), 3.93 (1H, br s), 3.73 (1H, br s), 3.14 (1H, br s),2.98 (1H, br s), 2.76 (1H, br s), 2.63 (1H, br s), 2.47 (1H, br s),2.02-1.90 (1H, m), 1.78-1.60 (3H, m), 1.46-1.35 (1H, m), 1.13 (4H, d,J=6.5 Hz). D2:N—((R)-1-((abs-1,5-cis)-6-azabicyclo[3.2.0]heptan-6-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide(4 mg). LCMS (ES+) 395 (M+H)⁺, RT 3.67 min (Analytical Method 2); ¹H NMR(400 MHz, DMSO) 8.31 (1H, d, J=8.2 Hz), 8.17 (2H, d, J=8.5 Hz), 8.04(2H, d, J=8.5 Hz), 3.94-3.86 (1H, m), 3.65 (1H, dd, J=4.7, 6.0 Hz), 3.18(1H, dd, J=7.9, 7.9 Hz), 2.93 (1H, dd, J=3.3, 7.5 Hz), 2.78-2.72 (1H,m), 2.61 (1H, dd, J=8.1, 11.3 Hz), 2.46 (1H, dd, J=5.9, 11.9 Hz),1.99-1.90 (1H, m), 1.73-1.56 (3H, m), 1.45-1.34 (1H, m), 1.17-1.13 (1H,m), 1.10 (3H, d, J=6.7 Hz).

Example 22:N-((2R)-1-(6,6-Dimethyl-3-azabicyclo[3.1.0]hexan-3-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Step 1:3-(2,4-Dimethoxybenzyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione

2,4-Dimethoxy benzylamine (1.45 g, 8.67 mmol) was added dropwise at 0°C. to a solution of 6,6-dimethyl-3-oxabicyclo[3.1.0]hexane-2,4-dione(1.21 g, 8.67 mmol) in THF (5 mL). The semi solid mixture was warmed to90° C. to remove the THF and the solid slowly melted as the mixture waswarmed to 180° C. for 2 h. The solution was cooled to 60° C. and pouredonto iced DIPE resulting in formation of a gum. This was separated andevaporated to afford the title compound as an orange gum (1.49 g, 59%).

Step 2: 3-(2,4-Dimethoxybenzyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane

Following method B from3-(2,4-dimethoxybenzyl)-3-azabicyclo[3.1.0]hexane-2,4-dione (1.49 g,5.15 mmol). The reaction was refluxed for 2 h and worked up in the usualway. This gave the title compound as a pale yellow gum (1.31 g). Theproduct was used in the next step without further purification.

Step 3: 6,6-Dimethyl-3-azabicyclo[3.1.0]hexane hydrochloride

3-(2,4-dimethoxybenzyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane (1.0 g,3.83 mmol) was dissolved in methanol (50 mL) and acetic acid (5 mL) then10% Pd/C (1.0 g) was added. The mixture was shaken under a hydrogen gasatmosphere (5 bar) for 18 h at r.t. The reaction mixture was thenfiltered through Celite, acidified with 4 N HCl in dioxane (5 mL) andconcentrated to yield the title compound as a pale pink solid (1.0 g)which was used in the next step without further purification.

Step 4: tert-Butyl((2R)-1-(6,6-dimethyl-3-azabicyclo[3.1.0]hexan-3-yl)-1-oxopropan-2-yl)carbamate

Following method A from (R)-2-((tert-butoxycarbonyl)amino)propanoic acid(659 mg, 3.48 mmol) and 6,6-dimethyl-3-azabicyclo[3.1.0]hexanehydrochloride (1.0 g, 3.83 mmol). The title compound was obtained as apale yellow gum (837 mg, 84%). The product was used in the next stepwithout further purification.

Step 5: tert-Butyl((2R)-1-(6,6-dimethyl-3-azabicyclo[3.1.0]hexan-3-yl)propan-2-yl)carbamate

Following method B from tert-butyl((2R)-1-(6,6-dimethyl-3-azabicyclo[3.1.0]hexan-3-yl)-1-oxopropan-2-yl)carbamate(837 mg, 2.96 mmol). The mixture was purified by passing through an SCXcartridge (elution with MeOH:7 M NH₃ in MeOH (9:1)). This gave the titlecompound as a pale yellow gum (525 mg, 66%).

Step 6:(2R)-1-(6,6-Dimethyl-3-azabicyclo[3.1.0]hexan-3-yl)propan-2-aminehydrochloride

Following method C from tert-butyl((2R)-1-(6,6-dimethyl-3-azabicyclo[3.1.0]hexan-3-yl)propan-2-yl)carbamate(525 mg, 1.96 mmol) gave the title compound as an orange solid (900mg, >100%). This was used in the next step without further purification.

Step 7:N-((2R)-1-(6,6-Dimethyl-3-azabicyclo[3.1.0]hexan-3-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide

Following method A from4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (459 mg, 1.78mmol) and(2R)-1-(6,6-dimethyl-3-azabicyclo[3.1.0]hexan-3-yl)propan-2-aminehydrochloride (900 mg, 1.95 mmol) and DIPEA (1.6 mL, 8.91 mmol) wereadded and the mixture stirred for 18 h. The mixture was washed withwater, passed through a phase separation cartridge and concentrated toyield the crude product. This residue was purified by preparative-HPLCto afford title compound as an off white solid (138 mg). LCMS (ES+) 409(M+H)⁺, RT 4.53 min (Analytical Method 1); ¹H NMR δ (ppm) (400 MHz,DMSO-d₆) 8.33 (1H, d, J=8.3 Hz), 8.15 (2H, d, J=8.7 Hz), 8.03 (2H, d,J=8.7 Hz), 4.13-4.04 (1H, m), 2.88 (2H, dd, J=9.2, 24.6 Hz), 2.63-2.54(3H, m), 2.40 (1H, dd, J=7.0, 11.7 Hz), 1.14-1.10 (8H, m), 0.92 (3H, s).

Example 23:(2S)-1-((R)-2-(3-Fluoro-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamido)propyl)-2-methylpyrrolidin-1-iumformate

Step 1: (Z)-3-Fluoro-4-(N-hydroxycarbamimidoyl)benzoic acid

To a stirred solution of 4-cyano-3-fluorobenzoic acid (2.00 g, 12.1mmol) in EtOH (30 mL) was added NH₂OH.HCl (1.18 g, 17.0 mmol) and KOH(2.03 g, 36.3 mmol). The mixture was stirred at r.t. for 17 h, thenneutralized with 1 M HCl_((aq)) and extracted into EtOAc (3×25 mL). Thecombined organics were dried (MgSO₄) and concentrated to give the titlecompound as a yellow oil which was progressed without furtherpurification.

Step 2: 3-Fluoro-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoicacid

To a stirred solution of (Z)-3-fluoro-4-(N-hydroxycarbamimidoyl)benzoicacid (12.1 mmol) in THF (30 mL) was added TFAA (2.50 mL, 18.2 mmol). Themixture was stirred for 3 h and then poured onto ice-water and acidifiedto pH 4 with 1 M HCl_((aq)), before extracting into EtOAc (3×30 mL). Thecombined organics were dried (MgSO₄) and concentrated. Purification byflash column chromatography (gradient elution i-hex [+3% AcOH] to 4:1i-hex:EtOAc [+3% AcOH]) gave the title compound as an off-white solid(150 mg, 4% (2 steps)). LCMS (ES+) 277 (M+H)⁺.

Step 3:(2S)-1-((R)-2-(3-Fluoro-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamido)propyl)-2-methylpyrrolidin-1-iumformate

Following method A from3-fluoro-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (75mg, 0.27 mmol) and intermediate 2(59 mg, 0.41 mmol). Purification bypreparative-HPLC gave the title compound as yellow solid (39 mg, 32%).LCMS (ES+) 447 (M+H)⁺, RT 2.74 min (Analytical method 1). ¹H NMR δ (ppm)(400 MHz, DMSO-d₆) 8.55 (1H, d, J=8.2 Hz), 8.21-8.17 (2H, m), 7.95-7.91(2H, m), 4.19-4.09 (1H, m), 3.13 (1H, ddd, J=4.1, 7.2, 8.9 Hz), 2.74(1H, dd, J=9.3, 11.7 Hz), 2.42-2.34 (1H, m), 2.28 (1H, dd, J=5.7, 11.3Hz), 2.20 (1H, q, J=8.7 Hz), 1.93-1.83 (1H, m), 1.73-1.63 (2H, m),1.39-1.25 (1H, m), 1.19 (3H, d, J=6.7 Hz), 1.05 (3H, d, J=6.0 Hz).

Example 24:N—((R)-1-((S)-2-Methylpyrrolidin-1-yl)propan-2-yl)-5-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)picolinamide

Step 1: (Z)-5-(N-Hydroxycarbamimidoyl)picolinic acid

To a stirred solution of 5-cyanopicolinic acid (1.51 g, 10.2 mmol) inEtOH (30 mL) was added NH₂OH.HCl (999 mg, 14.3 mmol) and KOH (1.71 g,30.6 mmol). The mixture was stirred at r.t. for 72 h, then neutralizedwith 1 M HCl_((aq)). The resulting precipitate was collected by vacuumfiltration, washed with H₂O and EtOH and dried under vacuum to give thetitle compound as a white solid (1.21 g, 63%).

Step 2: 5-(5-(Trifluoromethyl)-1,2,4-oxadiazol-3-yl)picolinic acid

To a stirred solution of (Z)-5-(N′-hydroxycarbamimidoyl)picolinic acid(1.21 g, 6.47 mmol) in THF (20 mL) was added TFAA (1.35 mL, 9.71 mmol).The mixture was stirred at r.t. for 17 h and then poured onto ice-waterand acidified to pH 4 with 1 M HCl_((aq)), before extracting into EtOAc(3×30 mL). The combined organics were dried (MgSO₄) and concentrated.The residue was taken-up in DCM/MeOH (10 mL, 1:1) and the insolublesolid collected by vacuum filtration to give the title compound as anoff-white solid (593 mg, 35%). LCMS (ES+) 260 (M+H)⁺.

Step 3:N—((R)-1-((S)-2-Methylpyrrolidin-1-yl)propan-2-yl)-5-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)picolinamide

Following method A from5-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)picolinic acid (200 mg, 0.77mmol) and intermediate 2 (166 mg, 1.16 mmol). Purification by reversephase chromatography (gradient elution, 5-95% MeCN in 0.1% formic acid)and then passage through SCX cartridge (elution with MeOH:7 M NH₃ inMeOH (9:1)) gave the title compound as a orange gum (72 mg, 24%). LCMS(ES+) 384 (M+H)⁺, RT 3.92 min (Analytical method 2). ¹H NMR δ (ppm) (400MHz, DMSO-d₆) 9.25 (1H, d, J=1.8 Hz), 8.73 (1H, d, J=8.7 Hz), 8.64 (1H,dd, J=2.1, 8.2 Hz), 8.26 (1H, d, J=8.2 Hz), 4.19-4.09 (1H, m), 3.15-3.07(1H, m), 2.82 (1H, dd, J=9.0, 11.8 Hz), 2.37-2.28 (1H, m), 2.23 (1H, dd,J=5.1, 10.9 Hz), 2.16 (1H, q, J=8.7 Hz), 1.91-1.80 (1H, m), 1.71-1.61(2H, m), 1.34-1.23 (1H, m), 1.21 (3H, d, J=6.5 Hz), 1.02 (3H, d, J=6.0Hz).

Table of examples Example Structure IUPAC Name  1

(2S)-2-Methyl-1-((R)-2-(3-methyl-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamido)propyl)pyrrolidin-1-ium formate  2

N-(2-(3-Azabicyclo[3.2.1]octan-3-yl)ethyl)-4-(5-(trifluoromethyl)-1,2,4- oxadiazol-3-yl)benzamide  3

(R)-N-(1-(Isoindolin-2-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol- 3-yl)benzamide  4

N-((2R)-1-(3-Azabicyclo[3.1.0]hexan-3-yl)propan-2-yl)-4-(5-(trifluoromethyl)- 1,2,4-oxadiazol-3-yl)benzamide 5

(S)-N-(1-Cyclopropyl-2-(pyrrolidin-1-yl)ethyl)-4-(5-(trifluoromethyl)-1,2,4- oxadiazol-3-yl)benzamide  6

(R)-N-(1-Cyclopropyl-2-(pyrrolidin-1-yl)ethyl)-4-(5-(trifluoromethyl)-1,2,4- oxadiazol-3-yl)benzamide  7

(R)-N-(1-(3,4-Dihydroisoquinolin- 2(1H)-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3- yl)benzamide  8

(R)-N-(1-(3-Phenylazetidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4- oxadiazol-3-yl)benzamide  9

D1: N-((R)-1-((abs)-3- (Difluoromethoxy)piperidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)- 1,2,4-oxadiazol-3-yl)benzamide10

D2: N-((R)-1-((abs)-3- (Difluoromethoxy)piperidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)- 1,2,4-oxadiazol-3-yl)benzamide11

(R)-N-(1-(5-Azaspiro[2.5]octan-5-yl)propan-2-yl)-4-(5-(trifluoromethyl)- 1,2,4-oxadiazol-3-yl)benzamide12

(R)-N-(1-(6-Azaspiro[2.5]octan-6-yl)propan-2-yl)-4-(5-(trifluoromethyl)- 1,2,4-oxadiazol-3-yl)benzamide13

(R)-N-(1-(Azepan-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3- yl)benzamide 14

(R)-N-(1-(1-Azaspiro[3.3]heptan-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)- 1,2,4-oxadiazol-3-yl)benzamide15

(R)-N-(1-(5-Azaspiro[2.4]heptan-5-yl)propan-2-yl)-4-(5-(trifluoromethyl)- 1,2,4-oxadiazol-3-yl)benzamide16

N-((R)-1-(3-Azabicyclo[3.2.1]octan-3-yl)propan-2-yl)-4-(5-(trifluoromethyl)- 1,2,4-oxadiazol-3-yl)benzamide17

2-((R)-2-(4-(5-(Trifluoromethyl)-1,2,4- oxadiazol-3-yl)benzamido)propyl)octahydrocyclopenta [c]pyrrol-2-ium formate 18

(R)-N-(1-(3,4-dihydro-2,7-naphthyridin- 2(1H)-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3- yl)benzamide 19

N-((2R)-1-(3-Azabicyclo[3.2.0]heptan- 3-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3- yl)benzamide 20

D1: N-((R)-1-((abs-1,5-cis)-6- Azabicyclo[3.2.0]heptan-6-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4- oxadiazol-3-yl)benzamide 21

D2: N-((R)-1-((abs-1,5-cis)-6- Azabicyclo[3.2.0]heptan-6-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4- oxadiazol-3-yl)benzamide 22

N-((2R)-1-(6,6-Dimethyl-3- azabicyclo[3.1.0]hexan-3-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4- oxadiazol-3-yl)benzamide 23

(2S)-1-((R)-2-(3-Fluoro-4-(5- (trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamido)propyl)-2- methylpyrrolidin-1-ium formate 24

N-((R)-1-((S)-2-Methylpyrrolidin-1-yl)propan-2-yl)-5-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)picolinamide 25

3-methyl-N-((R)-1-((S)-2- methylpyrrolidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3- yl)benzamide 26

N-((2R)-1- (hexahydrocyclopenta[c]pyrrol-2(1H)-yl)propan-2-yl)-4-(5-(trifluoromethyl)- 1,2,4-oxadiazol-3-yl)benzamide27

3-fluoro-N-((R)-1-((S)-2- methylpyrrolidin-1-yl)propan-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3- yl)benzamide

Example 25: Analysis of Inhibition of HDAC4 with Class IIa HistoneDeacetylase (HDAC) Inhibitors

The potency of Class IIa Histone Deacetylase (HDAC) inhibitors wasquantified by measuring the Histone Deacetylase 4 (HDAC4) catalyticdomain enzymatic activity using the fluorogenic substrate,Boc-Lys(TFA)-AMC. The substrate was deacetylated to Boc-Lys-AMC byHDAC4. Cleavage by trypsin resulted in the release of the fluorophoreAMC from the deacetylated substrate. The fluorescence of the sample wasdirectly related to the histone deacetylase activity in the sample.

Serially Dilute HDAC Inhibitor Compounds.

Serial dilutions of the HDAC inhibitors and control reference compound(1-(5-(3-((4-(1,3,4-oxadiazol-2-yl)phenoxy)methyl)-1,2,4-oxadiazol-5-yl)thiophen-2-yl)-2,2,2-trifluoroethanone)were made by first resuspending the lyophilized compound to a finalconcentration of 10 mM in 100% dimethyl sulfoxide (DMSO). Stocks of 60μL aliquots of the 10 mM compound in DMSO were prepared and stored at−20° C. From one stock aliquot of each tested compound and the referencecompound, a 16-point serial dilution was prepared according to Table 1using a 125 μL 16-channel Matrix multi-channel pipette (MatrixTechnologies Ltd).

TABLE 1 Serial Dilution of Compounds Concen- Dilu- Diluted tration tionSolutions Well (μM) ratio Volumes Concentration A 10000 — 60 μL 10 mMTest 1 compound/reference control Concentration B 5000 1:2 30 μL A + 30μL DMSO 2 Concentration C 2500 1:2 30 μL B + 30 μL DMSO 3 ConcentrationD 1000  1:2.5 30 μL C + 45 μL DMSO 4 Concentration E 500 1:2 30 μL D +30 μL DMSO 5 Concentration F 250 1:2  30 μL E + 30 μL DMSO 6Concentration G 125 1:2  30 μL F + 30 μL DMSO 7 Concentration H 62.5 1:230 μL G + 30 μL DMSO 8 Concentration I 31.25 1:2 30 μL H + 30 μL DMSO 9Concentration J 15.63 1:2  30 μL I + 30 μL DMSO 10 Concentration K 7.811:2  30 μL J + 30 μL DMSO 11 Concentration L 3.91 1:2 30 μL K + 30 μLDMSO 12 Concentration M 1.95 1:2  30 μL L + 30 μL DMSO 13 ConcentrationN 0.98 1:2 30 μL M + 30 μL DMSO  14 Concentration O 0.49 1:2 30 μL N +30 μL DMSO 15 Concentration P 0.24 1:2 30 μL O + 30 μL DMSO 16

2 μL (200×) of each diluted solution and each control (full activity:100% DMSO alone or full inhibition 1 mM) was stamped into V-bottomedpolypropylene 384-well compound plates using either the Bravo (384-wellhead from Agilent) or 12.5 μL 16-channel Matrix multi-channel pipette(Matrix Technologies Ltd). Each well with the 200× compound solution wasdiluted 1:20 by the addition of 38 μL assay buffer+DMSO (10.5% DMSO, 45mM Tris-HCl, 123 mM NaCl, 2.4 mM KCl, and 0.9 mM MgCl₂ at pH 8.0 andequilibrated to r.t.).

Prepare HDAC4 Catalytic Domain Enzyme (0.2 μg/mL).

The HDAC4 catalytic domain enzyme was human catalytic domain HDAC4protein (amino acids 648-1032) with a C-terminal 6× histidine tag,produced by BioFocus. A working solution of enzyme was prepared from a500 g/mL stock aliquot of HDAC4 catalytic domain (thawed on ice) dilutedto 0.2 μg/mL with assay buffer (50 mM Tris-HCl, 137 mM NaCl, 2.7 mM KCl,and 1 mM MgCl₂ at pH 8 and equilibrated to r.t.) just prior to theaddition of the enzyme to the assay.

Prepare 5× (50 μM) Boc-Lys(TFA)-AMC Substrate.

5× (50 μM) substrate was prepared just prior to the addition to theassay. A 1 mM substrate stock was made by diluting a 100 mMBoc-Lys(TFA)-AMC in DMSO solution 1:100 by adding it drop-wise to assaybuffer (equilibrated to r.t.) while vortexing at slow speed to preventprecipitation. The 5× substrate was prepared by diluting the 1 mMsubstrate solution 1:20 by adding it drop-wise to assay buffer(equilibrated to r.t.) while vortexing at slow speed to preventprecipitation.

Prepare 3× (30 μM) Developer/Stop Solution.

3× (30 μM) Developer/Stop Solution was prepared just prior to additionto the plate by diluting a stock solution of 10 mM reference compound1:333 in 25 mg/mL trypsin (PAA Laboratories Ltd.) equilibrated to r.t.

Assay.

5 μL of each solution of 1:20 diluted compound from above wastransferred to a clear bottomed, black, 384-well assay plate using theBravo or the Janus (384-well MDT head from Perkin Elmer). Using a16-channel Matrix multi-channel pipette, 35 μL of the working solutionof HDAC4 catalytic domain enzyme (0.2 μg/mL in assay buffer) wastransferred to the assay plate. The assay was then started by adding 10μL of 5× (50 μM) substrate to the assay plates using either the Bravo,Janus or 16-channel Matrix multi-channel pipette. The assay plate wasthen shaken for two minutes on an orbital shaker at 900 rpm (rotationsper minute). Next the plate was incubated for 15 minutes at 37° C. Thereaction was stopped by adding 25 μL of 3× (30 μM) developer/stopsolution to the assay plates using either the Bravo, Janus or a16-channel Matrix multi-channel pipette. Assay plates were then shakenfor 5 minutes on an orbital shaker at 1200 rpm. Next, the assay plateswere incubated at 37° C. for 1 hour in a tissue culture incubator.Finally, the fluorescence was measured (Excitation: 355 nm, Emission:460 nm) using PerkinElmer EnVision in top read mode.

Example 26: Analysis of Inhibition of HDAC5 with Class Ha HistoneDeacetylase (HDAC) Inhibitors

The potency of Class IIa Histone Deacetylase (HDAC) inhibitors isquantified by measuring the Histone Deacetylase 5 (HDAC5) enzymaticactivity using the fluorogenic substrate, Boc-Lys(TFA)-AMC. Thesubstrate is deacetylated to Boc-Lys-AMC by HDAC5. Cleavage by trypsinresults in the release of the fluorophore AMC from the deacetylatedsubstrate. The fluorescence of the sample is directly related to thehistone deacetylase activity in the sample.

Serially Dilute HDAC Inhibitor Compounds.

Serial dilutions of the HDAC inhibitors and control reference compound(1-(5-(3-((4-(1,3,4-oxadiazol-2-yl)phenoxy)methyl)-1,2,4-oxadiazol-5-yl)thiophen-2-yl)-2,2,2-trifluoroethanone)are made by first resuspending the lyophilized compound to a finalconcentration of 10 mM in 100% DMSO. Stocks of 60 μL aliquots of the 10mM compound in DMSO are prepared and stored at −20° C. From one stockaliquot of each compound to be tested and the reference compound, a16-point serial dilution is prepared according to Table 1 using a 125 μL16-channel Matrix multi-channel pipette.

2 μL (200×) of each diluted solution and each control (full activity:100% DMSO alone or full inhibition 1 mM) is stamped into V-bottompolypropylene 384-well compound plates using either Bravo, Janus, or a12.5 μL 16-channel Matrix multi-channel pipette. Each well with the 2 μLof the 200× stamped compound solution is diluted 1:20 by the addition of38 μL assay buffer+DMSO (10.5% DMSO, 45 mM Tris-HCl, 123 mM NaCl, 2.4 mMKCl, and 0.9 mM MgCl₂ at pH 8.0 and equilibrated to 37° C.).

Prepare HDAC5 Catalytic Domain Enzyme (0.57 μg/mL).

The HDAC5 catalytic domain enzyme is human HDAC5 catalytic domain(GenBank Accession No. NM_001015053), amino acids 657-1123 with aC-terminal His tag and can be obtained from BPS BioScience. The proteinis 51 kDa and is expressed in a baculovirus expression system. A workingsolution of enzyme is prepared from a 1.65 mg/mL stock aliquot of HDAC5catalytic domain (thawed on ice) diluted to 0.57 μg/mL with assay buffer(50 mM Tris-HCl, 137 mM NaCl, 2.7 mM KCl, and 1 mM MgCl₂ at pH 8 andequilibrated to 37° C.) just prior to the addition of the enzyme to theassay.

Prepare 5× (40 μM) Boc-Lys(TFA)-AMC Substrate.

5× (40 μM) substrate is prepared just prior to the addition to theassay. The 5× substrate is prepared by diluting the 100 mMBoc-Lys(TFA)-AMC in DMSO solution 1:2500 by adding it drop-wise to assaybuffer (equilibrated to 37° C.) while vortexing at slow speed to preventprecipitation.

Prepare 3× (30 μM) Developer/Stop Solution.

3× (30 μM) Developer/Stop Solution is prepared just prior to addition tothe plate by diluting a stock solution of 10 mM reference compound 1:333in 25 mg/mL trypsin equilibrated to 37° C.

Assay.

5 μL of each solution of the 1:20 diluted inhibitor compounds andcontrols from above is transferred to a clear bottomed, black, 384-wellassay plate using the Bravo or Janus. Using a 16-channel Matrixmulti-channel pipette, 35 μL of the working solution of the HDAC5catalytic domain enzyme (0.57 μg/mL in assay buffer) is transferred tothe assay plate. The assay is then started by adding 10 μL of 5× (40 μM)substrate to the assay plates using either the Bravo, Janus or16-channel Matrix multi-channel pipette. The assay plate is then shakenfor one minute on an orbital shaker at 900 rpm. Next, the plates areincubated for 15 minutes at 37° C. The reaction is stopped by adding 25μL of 3× (30 μM) developer/stop solution to the assay plates usingeither the Bravo, Janus or a 16-channel Matrix multi-channel pipette.Assay plates are then shaken for 2 minutes on an orbital shaker at 900rpm. Next, the assay plates are incubated at 37° C. for 1 hour in atissue culture incubator followed by shaking for 1 minute at the maximumrpm on an orbital shaker before reading on the EnVision. Finally, thefluorescence is measured (Excitation: 355 nm, Emission: 460 nm) usingPerkinElmer EnVision in top read mode.

Example 27: Analysis of Inhibition of HDAC7 with Class IIa HistoneDeacetylase (HDAC) Inhibitors

The potency of Class IIa Histone Deacetylase (HDAC) inhibitors isquantified by measuring the Histone Deacetylase 7 (HDAC7) enzymaticactivity using the fluorogenic substrate, Boc-Lys(TFA)-AMC. Thesubstrate is deacetylated to Boc-Lys-AMC by HDAC7. Cleavage by trypsinresults in the release of the fluorophore AMC from the deacetylatedsubstrate. The fluorescence of the sample is directly related to thehistone deacetylase activity in the sample.

Serially Dilute HDAC Inhibitor Compounds.

Serial dilutions of the HDAC inhibitors and control reference compound(1-(5-(3-((4-(1,3,4-oxadiazol-2-yl)phenoxy)methyl)-1,2,4-oxadiazol-5-yl)thiophen-2-yl)-2,2,2-trifluoroethanone)are made by first resuspending the lyophilized compound to a finalconcentration of 10 mM in 100% DMSO. Stocks of 60 μL aliquots of the 10mM compound in DMSO are prepared and stored at −20° C. From one stockaliquot of each compound to be tested and the reference compound, a16-point serial dilution is prepared according to Table 1 using a 125 μL16-channel Matrix multi-channel pipette.

2 μL (200×) of each diluted solution and each control (full activity:100% DMSO alone or full inhibition 1 mM) is stamped into V-bottompolypropylene 384-well compound plates using either the Bravo, Janus, ora 12.5 μL 16-channel Matrix multi-channel pipette. Each well with the200× compound solution is diluted 1:20 by the addition of 38 μL assaybuffer+DMSO (10.5% DMSO, 45 mM Tris-HCl, 123 mM NaCl, 2.4 mM KCl, and0.9 mM MgCl₂ at pH 8.0 and equilibrated to 37° C.).

Prepare HDAC7 Enzyme (71 ng/mL).

The HDAC7 enzyme is human HDAC7 (GenBank Accession No. AY302468) aminoacids 518-end with a N-terminal Glutathione S-transferase (GST) tag andcan be obtained from BPS BioScience. The protein is 78 kDa and isexpressed in a baculovirus expression system. A working solution ofenzyme is prepared from a 0.5 mg/mL stock aliquot of HDAC7 (thawed onice) diluted to 71 ng/mL with assay buffer (50 mM Tris-HCl, 137 mM NaCl,2.7 mM KCl, and 1 mM MgCl₂ at pH 8 and equilibrated to 37° C.) justprior to the addition of enzyme to the assay.

Prepare 5× (50 μM)Boc-Lys(TFA)-AMC Substrate.

5× (50 μM) substrate is prepared just prior to the addition to theassay. The 5× substrate is prepared by diluting a 100 mMBoc-Lys(TFA)-AMC in DMSO solution 1:2000 by adding it drop-wise to assaybuffer (equilibrated to 37° C.) while vortexing at slow speed to preventprecipitation.

Prepare 3× (30 μM) Developer/Stop Solution.

3× (30 μM) Developer/Stop Solution is prepared just prior to addition tothe plate by diluting a stock solution of 10 mM reference compound 1:333in 25 mg/mL trypsin equilibrated to 37° C.

Assay.

5 μL of each solution of 1:20 diluted compound from above is transferredto a clear bottomed, black, 384-well assay plate using the Bravo orJanus. Using a 16-channel Matrix multi-channel pipette, 35 μL of theworking solution of the HDAC7 enzyme (71 ng/mL in assay buffer) istransferred to the assay plate. The assay is then started by adding 10μL of 5× (50 μM) substrate to the assay plate using either the Bravo,Janus or 16-channel Matrix multi-channel pipette. The assay plate isthen shaken for one minute on an orbital shaker at 900 rpm. Next, theplate is incubated for 15 minutes at 37° C. The reaction is then stoppedby adding 25 μL of 3× (30 μM) developer/stop solution to the assayplates using either the Bravo, Janus or a 16-channel Matrixmulti-channel pipette. The assay plate is then shaken for 2 minutes onan orbital shaker at 900 rpm. Next, the assay plate is incubated at 37°C. for 1 hour in a tissue culture incubator followed by shaking for 1minute at maximum rpm on an orbital shaker. Finally, the fluorescence ismeasured (Excitation: 355 nm, Emission: 460 nm) using PerkinElmerEnVision in top read mode.

Example 28: Analysis of Inhibition of HDAC9 with Class IIa HistoneDeacetylase (HDAC) Inhibitors

The potency of Class IIa Histone Deacetylase (HDAC) inhibitors isquantified by measuring the Histone Deacetylase 9 (HDAC9) enzymaticactivity using the fluorogenic substrate, Boc-Lys(TFA)-AMC. Thesubstrate is deacetylated to Boc-Lys-AMC by HDAC9. Cleavage by trypsinresults in the release of the fluorophore AMC from the deacetylatedsubstrate. The fluorescence of the sample is directly related to thehistone deacetylase activity in the sample.

Serially Dilute HDAC Inhibitor Compounds.

Serial dilutions of the HDAC inhibitors and control reference compound(1-(5-(3-((4-(1,3,4-oxadiazol-2-yl)phenoxy)methyl)-1,2,4-oxadiazol-5-yl)thiophen-2-yl)-2,2,2-trifluoroethanone)are made by first resuspending the lyophilized compound to a finalconcentration of 10 mM in 100% DMSO. Stocks of 60 μL aliquots of the 10mM compound in DMSO are prepared and stored at −20° C. From one stockaliquot of each compound to be tested and the reference compound, a16-point serial dilution is prepared according to Table 1 using a 125 μL16-channel Matrix multi-channel pipette.

2 μL (200×) of each diluted solution and each control (full activity:100% DMSO alone or full inhibition 1 mM) is stamped into V-bottompolypropylene 384-well compound plates using either the Bravo, Janus, or12.5 μL 16-channel Matrix multi-channel pipette. Each well with thestamped 200× compound solution is diluted 1:20 by the addition of 38 μLassay buffer+DMSO (10.5% DMSO, 45 mM Tris-HCl, 123 mM NaCl, 2.4 mM KCl,and 0.9 mM MgCl₂ at pH 8.0 and equilibrated to 37° C.).

Prepare HDAC9 Enzyme (0.57 μg/mL).

The HDAC9 enzyme is human HDAC9 (GenBank Accession No. NM_178423) aminoacids 604-1066 with a C-terminal His tag and can be obtained from BPSBioScience. The protein is 50.7 kDa and is expressed in a baculovirusexpression system. A working solution of enzyme is prepared from a 0.5mg/mL stock aliquot of HDAC9 (thawed on ice) diluted to 0.57 μg/mL withassay buffer (50 mM Tris-HCl, 137 mM NaCl, 2.7 mM KCl, and 1 mM MgCl₂ atpH 8 and equilibrated to 37° C.) just prior to the addition of enzyme tothe assay.

Prepare 5× (125 μM) Boc-Lys(TFA)-AMC Substrate.

5× (125 μM) substrate is prepared just prior to the addition to theassay. The 5× substrate is prepared by diluting a 100 mMBoc-Lys(TFA)-AMC in DMSO solution 1:800 by adding it drop-wise to assaybuffer (equilibrated to 37° C.) while vortexing at slow speed to preventprecipitation.

Prepare 3× (30 μM) Developer/Stop Solution.

3× (30 μM) Developer/Stop Solution is prepared just prior to addition tothe plate by diluting a stock solution of 10 mM reference compound 1:333in 25 mg/mL trypsin equilibrated to 37° C.

Assay.

5 μL of each solution of 1:20 diluted compound from above is transferredto a clear bottomed, black, 384-well assay plate using the Bravo orJanus. Using a 16-channel Matrix multi-channel pipette, 35 μL of theworking solution of the HDAC9 enzyme (0.57 μg/mL in assay buffer) istransferred to the assay plate. The assay is then started by adding 10μL of 5× (125 μM) substrate to the assay plate using either the Bravo,Janus or 16-channel Matrix multi-channel pipette. The assay plate isthen shaken for one minute on an orbital shaker at 900 rpm. Next, theplate is incubated for 15 minutes at 37° C. The reaction is stopped byadding 25 μL of 3× developer/stop solution to the assay plates usingeither the Bravo, Janus or a 16-channel Matrix multi-channel pipette.The assay plate is then shaken for 2 minutes on an orbital shaker at 900rpm. Next, the assay plate is incubated at 37° C. for 1 hour in a tissueculture incubator followed by shaking for 1 minute at maximum rpm on anorbital shaker before reading on the enVision. Finally, the fluorescenceis measured (Excitation: 355 nm, Emission: 460 nm) using PerkinElmerEnVision in top read mode.

Example 29: Analysis of Inhibition of Cellular Class IIa HDAC Activitywith Class IIa Histone Deacetylase (HDAC) Inhibitors; Cell (Lys-TFA)Substrate

The potency of Class IIa Histone Deacetylase (HDAC) inhibitors wasquantified by measuring the cellular histone deacetylase enzymaticactivity using the fluorogenic substrate, Boc-Lys(TFA)-AMC. Afterpenetration in Jurkat E6-1 cells, the substrate was deacetylated toBoc-Lys-AMC. After cell lysis and cleavage by trypsin, the fluorophoreAMC was released from the deacetylated substrate only. The fluorescenceof the sample was directly related to the histone deacetylase activityin the sample.

Jurkat E6.1 Cell Culture and Plating.

Jurkat E6.1 cells were cultured according to standard cell cultureprotocols in Jurkat E6.1 Growth Media (RPMI without phenol red, 10% FBS,10 mM HEPES, and 1 mM Sodium Pyruvate). Jurkat E6.1 cells were countedusing a Coulter Counter and resuspended in Jurkat E6.1 growth media at aconcentration of 75,000 cells/35 μL. 35 μL or 75,000 cells was seededinto Greiner microtitre assay plates. The plates were then incubated at37° C. and 5% CO₂ while other assay components were being prepared.

Serially Dilute HDAC Inhibitor Compounds.

Serial dilutions of the HDAC inhibitors and control reference compound(1-(5-(3-((4-(1,3,4-oxadiazol-2-yl)phenoxy)methyl)-1,2,4-oxadiazol-5-yl)thiophen-2-yl)-2,2,2-trifluoroethanone)were made by first resuspending the lyophilized compound to a finalconcentration of 10 mM in 100% DMSO. Stocks of 70 μL aliquots of the 10mM compound in DMSO were prepared and stored at −20° C. From one stockaliquot of each tested compound and the reference compound, a 16-pointserial dilution was prepared according to Table 1 using a 125 μL16-channel Matrix multi-channel pipette.

2 μL (200×) of each diluted solution and each control (full activity:100% DMSO alone or full inhibition 1 mM) was stamped into V-bottompolypropylene 384-well compound plates using either the Bravo, Janus, or12.5 μL 16-channel Matrix multi-channel pipette. Each well with the 200×compound solution was diluted 1:20 by the addition of 38 μL Jurkat assaybuffer+DMSO (9.5% DMSO, RPMI without phenol red, 0.09% FBS, 9 mM Hepes,and 0.9 mM Sodium Pyruvate equilibrated to r.t.)

Prepare 5× (500 μM) Boc-Lys(TFA)-AMC Substrate.

5× (500 μM) substrate was prepared just prior to the addition to theassay. The 5× substrate was prepared by diluting a 100 mMBoc-Lys(TFA)-AMC in DMSO solution 1:200 by adding it drop-wise to Jurkatassay medium (RPMI without phenol red, 0.1% FBS, 10 mM Hepes, and 1 mMSodium Pyruvate equilibrated to 37° C.) while vortexing at slow speed toprevent precipitation.

Prepare 3× Lysis Buffer.

10 mL of 3× lysis buffer was prepared with 8.8 mL of 3× stock lysisbuffer (50 mM Tris-HCl, pH 8.0, 137 mM NaCl, 2.7 mM KCl, 1 mM MgCl₂, 1%Nonidet P40 Substitute equilibrated to r.t.) and 1.2 mL of 3 mg/mLTrypsin equilibrated to r.t.

Assay.

5 μL of each solution of 1:20 diluted compound from above wastransferred to the Greiner microtitre assay plates with 75,000cells/well using the Bravo. Cells were then incubated for 2 hours at 37°C. and 5% CO₂. The assay was then started by adding 10 μL of 5× (500 μM)substrate to the assay plate using either the Bravo or 16-channel Matrixmulti-channel pipette. The cells were then incubated for 3 hours at 37°C. and 5% CO₂. Next, 25 μL of 3× lysis buffer was added to each wellusing either the 125 μL 16 channel pipette or the Bravo. The assay platewas then incubated overnight (15-16 hours) at 37° C. and 5% CO₂. Thefollowing day, the plates were shaken on an orbital shaker for 1 minuteat 900 rpm. Finally the top read fluorescence (Excitation: 355 nm,Emission: 460 nm) was measured using PerkinElmer EnVision.

Example 30: Analysis of Inhibition of Cellular Class I HDAC Activitywith Class IIa Histone Deacetylase (HDAC) Inhibitors; Cell (Lys-Ac)Substrate

The Class I HDAC activity of Class IIa Histone Deacetylase (HDAC)inhibitors was quantified by measuring the cellular histone deacetylaseenzymatic activity using the fluorogenic substrate, Boc-Lys(Ac)-AMC.This was performed according to the procedure in Example 29, usingBoc-Lys(Ac)-AMC substrate in place of Boc-Lys(TFA)-AMC.

Using the synthetic methods similar to those described above and theassay protocols described above, the following compounds weresynthesized and tested. Examples 1, 17, and 23 in the table below areshown as a formate salt, but it is contemplated that the free base wouldperform in an equivalent manner in the assays.

HDAC4 Cell Cell Biochemical (Lys-TFA) (Lys-Ac) Example Structure IC₅₀(μM) IC₅₀ (μM) IC₅₀ (μM)  1

0.10 1.6 >50  2

0.14 0.33 4.9  3

0.039 0.14 4.9  4

0.008 0.051 6.9  5

0.048 0.71 25.7  6

0.077 0.75 >50  7

0.009 0.018 2.4  8

0.014 0.039 0.71  9

0.021 0.057 3.7 10

0.003 0.007 1.5 11

0.003 0.009 0.97 12

0.030 0.058 4.0 13

0.011 0.030 1.7 14

0.016 0.18 7.7 15

0.008 0.069 5.1 16

0.007 0.012 0.89 17

0.012 0.028 0.69 18

0.006 0.19 12.7 19

0.004 0.009 1.1 20

0.022 0.15 7.8 21

0.004 0.016 3.1 22

0.020 0.086 6.1 23

0.009 0.072 21.4 24

0.134 1.03 33.7

REFERENCES

-   1) Bioorganic & Medicinal Chemistry Letters 17 (2007) 6476-6480

While some embodiments have been shown and described, variousmodifications and substitutions may be made thereto without departingfrom the spirit and scope of the disclosure. For example, for claimconstruction purposes, it is not intended that the claims set forthhereinafter be construed in any way narrower than the literal languagethereof, and it is thus not intended that exemplary embodiments from thespecification be read into the claims. Accordingly, it is to beunderstood that the present disclosure has been described by way ofillustration and not limitations on the scope of the claims.

What is claimed:
 1. A compound of Formula I:

or a pharmaceutically acceptable salt, an optical isomer, or a mixtureof optical isomers thereof; wherein: W is N or CR⁵; X is N or CR⁶; Y isN or CR⁷; and Z is N or CR⁸; provided not more than two of W, X, Y, andZ are N; R¹ is selected from H and C₁-C₃ alkyl; R² is C₂-C₃ alkyleneoptionally substituted with C₁-C₂ haloalkyl or 3 or 4-memberedcycloalkyl; R³ and R⁴, together with the nitrogen atom to which they areattached, form: a 4 or 7-membered heteromonocyclic group optionallysubstituted with one to five substituents independently selected from:C₁-C₃ alkoxy, C₁-C₃ alkyl, C₁-C₃ haloalkoxy, C₁-C₃ haloalkyl, 3 or4-membered cycloalkoxy, 3 or 4-membered cycloalkyl, 3 or 4-memberedheterocycloalkyl, carboxy, aryl, cyano, halo, and heteroaryl, whereinaryl and heteroaryl are optionally further substituted with one to fivesubstituents independently selected from C₁-C₃ alkyl, C₁-C₃ haloalkyl,and halo; or a 5 or 6-membered heteromonocyclic group optionallysubstituted with one to five substituents independently selected from:C₁-C₃ alkoxy, C₁-C₃ alkyl, C₁-C₃ haloalkoxy, C₁-C₃ haloalkyl, 3 or4-membered cycloalkoxy, 3 or 4-membered cycloalkyl, 3 or 4-memberedheterocycloalkyl, carboxy, aryl, cyano, halo, and heteroaryl, whereinaryl and heteroaryl are optionally further substituted with one to fivesubstituents each independently selected from C₁-C₃ alkyl, C₁-C₃haloalkyl, and halo; or a 6, 7, 8, 9, or 10-membered heterobicyclicgroup, which is optionally substituted with one to five substituentsindependently selected from: C₁-C₃ alkoxy, C₁-C₃ alkyl, C₁-C₃haloalkoxy, C₁-C₃ haloalkyl, 3 or 4-membered cycloalkoxy, 3 or4-membered cycloalkyl, 3 or 4-membered heterocycloalkyl, carboxy, aryl,cyano, halo, and heteroaryl, wherein aryl and heteroaryl are optionallyfurther substituted with one to five substituents independently selectedfrom C₁-C₃ alkyl, C₁-C₃ haloalkyl, and halo; R⁵, R⁶, R⁷ and R⁸ are eachindependently selected from H, C₁-C₄ alkyl, C₁-C₄ haloalkyl, and halo;provided that if R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 5 or 6-membered heteromonocyclic group, then:R² is C₂-C₃ alkylene substituted with C₁-C₂ haloalkyl or 3 or 4-memberedcycloalkyl.
 2. A compound according to claim 1, or a pharmaceuticallyacceptable salt, an optical isomer, or a mixture of optical isomersthereof, wherein the compound is of Formula II:


3. A compound according to claim 1, or a pharmaceutically acceptablesalt, an optical isomer, or a mixture of optical isomers thereof,wherein R² is C₂-C₃ alkylene optionally substituted with 3 or 4-memberedcycloalkyl.
 4. A compound of Formula III:

or a pharmaceutically acceptable salt, an optical isomer, or a mixtureof optical isomers thereof, wherein: R¹, R⁵, R⁶, R⁷ and R⁸ are each H;and R³ and R⁴, together with the nitrogen atom to which they areattached, form a 5 or 6-membered heteromonocyclic group substituted withone to two substituents each independently selected from 3 or 4-memberedcycloalkyl and heteroaryl.
 5. A compound according to claim 1, or apharmaceutically acceptable salt, an optical isomer, or a mixture ofoptical isomers thereof, wherein R⁶ is H.
 6. A compound according toclaim 1, or a pharmaceutically acceptable salt, an optical isomer, or amixture of optical isomers thereof, wherein the compound is of FormulaV:


7. A compound according to claim 6, or a pharmaceutically acceptablesalt, an optical isomer, or a mixture of optical isomers thereof,wherein R² is C₂-C₃ alkylene optionally substituted with 3 or 4-memberedcycloalkyl.
 8. A compound according to claim 7, or a pharmaceuticallyacceptable salt, an optical isomer, or a mixture of optical isomersthereof, wherein the compound is of Formula VI:


9. A compound according to claim 1, or a pharmaceutically acceptablesalt, an optical isomer, or a mixture of optical isomers thereof,wherein R⁵ is H.
 10. A compound according to claim 1, or apharmaceutically acceptable salt, an optical isomer, or a mixture ofoptical isomers thereof, wherein R⁷ is H.
 11. A compound according toclaim 1, or a pharmaceutically acceptable salt, an optical isomer, or amixture of optical isomers thereof, wherein R⁸ is selected from H, halo,and methyl.
 12. A compound according to claim 1, or a pharmaceuticallyacceptable salt, an optical isomer, or a mixture of optical isomersthereof, wherein R¹ is selected from H and methyl.
 13. A compoundaccording to claim 12, or a pharmaceutically acceptable salt, an opticalisomer, or a mixture of optical isomers thereof, wherein R¹ is H.
 14. Acompound according to claim 1, or a pharmaceutically acceptable salt, anoptical isomer, or a mixture of optical isomers thereof, wherein R³ andR⁴, together with the nitrogen atom to which they are attached, form a 4or 7-membered heteromonocyclic group selected from3-phenylazetidin-1-yl, and azepan-1-yl.
 15. A compound according toclaim 1, or a pharmaceutically acceptable salt, an optical isomer, or amixture of optical isomers thereof, wherein R³ and R⁴, together with thenitrogen atom to which they are attached, form a 5 or 6-memberedheteromonocyclic group optionally substituted with one to fivesubstituents independently selected from: C₁-C₃ alkoxy, C₁-C₃ alkyl,C₁-C₃ haloalkoxy, C₁-C₃ haloalkyl, 3 or 4-membered cycloalkoxy, 3 or4-membered cycloalkyl, 3 or 4-membered heterocycloalkyl, aryl, andheteroaryl, wherein aryl and heteroaryl are optionally furthersubstituted with one to five substituents independently selected fromC₁-C₃ alkyl, C₁-C₃ haloalkyl, and halo.
 16. A compound according toclaim 1, or a pharmaceutically acceptable salt, an optical isomer, or amixture of optical isomers thereof, wherein R³ and R⁴, together with thenitrogen atom to which they are attached, form a 5 or 6-memberedheteromonocyclic group optionally substituted with one to fivesubstituents independently selected from: C₁-C₃ alkyl, C₁-C₃ haloalkoxy,and aryl, wherein aryl is optionally further substituted with one tofive substituents independently selected from C₁-C₃ alkyl, C₁-C₃haloalkyl, and halo.
 17. A compound according to claim 1, or apharmaceutically acceptable salt, an optical isomer, or a mixture ofoptical isomers thereof, wherein R³ and R⁴, together with the nitrogenatom to which they are attached, form a 5 or 6-membered heteromonocyclicgroup optionally substituted with one to five substituents independentlyselected from: C₁-C₃ alkyl, C₁-C₃ haloalkoxy, and phenyl, wherein phenylis optionally further substituted with one to five substituentsindependently selected from C₁-C₃ alkyl, C₁-C₃ haloalkyl, and halo. 18.A compound according to claim 1, or a pharmaceutically acceptable salt,an optical isomer, or a mixture of optical isomers thereof, wherein R³and R⁴, together with the nitrogen atom to which they are attached, forma 5 or 6-membered heteromonocyclic group optionally substituted with oneto five substituents independently selected from: methyl,difluoromethoxy, and phenyl.
 19. A compound according to claim 1, or apharmaceutically acceptable salt, an optical isomer, or a mixture ofoptical isomers thereof, wherein R³ and R⁴, together with the nitrogenatom to which they are attached, form a 5 or 6-membered heteromonocyclicgroup selected from pyrrolidin-1-yl, and piperidin-1-yl, each of whichis optionally substituted with one substituent selected from: methyl,difluoromethoxy, and phenyl.
 20. A compound according to claim 1, or apharmaceutically acceptable salt, an optical isomer, or a mixture ofoptical isomers thereof, wherein R³ and R⁴, together with the nitrogenatom to which they are attached, form a 5 or 6-membered heteromonocyclicgroup selected from pyrrolidin-1-yl, 2-methyl-pyrrolidin-1-yl, and3-(difluoromethoxy)piperidin-1-yl.
 21. A compound according to claim 4,or a pharmaceutically acceptable salt, an optical isomer, or a mixtureof optical isomers therof, wherein R³ and R⁴, together with the nitrogenatom to which they are attached, form a 5-membered heteromonocyclicgroup substituted with one 3 or 4-membered cycloalkyl.
 22. A compoundaccording to claim 4, or a pharmaceutically acceptable salt, an opticalisomer, or a mixture of optical isomers therof, wherein R³ and R⁴,together with the nitrogen atom to which they are attached, form a5-membered heteromonocyclic group substituted with one heteroaryl.
 23. Acompound according to claim 1, or a pharmaceutically acceptable salt, anoptical isomer, or a mixture of optical isomers thereof, wherein R⁵ isC₁-C₄ alkyl or halo.
 24. A compound according to claim 1, or apharmaceutically acceptable salt, an optical isomer, or a mixture ofoptical isomers thereof, wherein R⁸ is C₁-C₄ alkyl or halo.
 25. Acompound according to claim 1, or a pharmaceutically acceptable salt, anoptical isomer, or a mixture of optical isomers thereof, wherein W is N.26. A compound according to claim 1, or a pharmaceutically acceptablesalt, an optical isomer, or a mixture of optical isomers thereof,wherein Z is N.
 27. A compound according to claim 1, or apharmaceutically acceptable salt, an optical isomer, or a mixture ofoptical isomers thereof, wherein R³ and R⁴, together with the nitrogenatom to which they are attached, form a 6, 7, or 8-memberedheterobicyclic group, which is optionally substituted with one to fivesubstituents independently selected from: C₁-C₃ alkoxy, C₁-C₃ alkyl,C₁-C₃ haloalkoxy, C₁-C₃ haloalkyl, 3 or 4-membered cycloalkoxy, 3 or4-membered cycloalkyl, 3 or 4-membered heterocycloalkyl, carboxy, aryl,cyano, halo, and heteroaryl, wherein aryl and heteroaryl are optionallyfurther substituted with one to five substituents independently selectedfrom C₁-C₃ alkyl, C₁-C₃ haloalkyl, and halo.
 28. A compound according toclaim 1, or a pharmaceutically acceptable salt, an optical isomer, or amixture of optical isomers thereof, wherein R³ and R⁴, together with thenitrogen atom to which they are attached, form a 6, 7, 8, 9, or10-membered heterobicyclic group selected from:1-azaspiro[3.3]heptan-1-yl, 3-azabicyclo[3.1.0]hexan-3-yl,3-azabicyclo[3.2.0]heptan-3-yl, 3-azabicyclo[3.2.1]heptan-3-yl,3-azabicyclo[3.2.1]octan-3-yl, 3,4-dihydroisoquinolin-2(1H)-yl,3,4-dihydro-2,7-naphthyridin-2(1H)-yl, 5-azaspiro[2.4]heptan-5-yl,5-azaspiro[2.5]octan-5-yl, 6-azaspiro[2.5]octan-6-yl,1-azaspiro[3.3]heptan-1-yl, 6-azabicyclo[3.2.0]heptan-6-yl,isoindolin-2-yl, and octahydrocyclopenta[c]pyrrol-2-yl, each of which isoptionally substituted with one to five substituents independentlyselected from: C₁-C₃ alkoxy, C₁-C₃ alkyl, C₁-C₃ haloalkoxy, C₁-C₃haloalkyl, 3 or 4-membered cycloalkoxy, 3 or 4-membered cycloalkyl, 3 or4-membered heterocycloalkyl, carboxy, aryl, cyano, halo, and heteroaryl,wherein aryl and heteroaryl are optionally further substituted with oneto five substituents independently selected from C₁-C₃ alkyl, C₁-C₃haloalkyl, and halo.
 29. A compound according to claim 1, or apharmaceutically acceptable salt, an optical isomer, or a mixture ofoptical isomers thereof, wherein R³ and R⁴, together with the nitrogenatom to which they are attached, form a 6, 7, 8, 9, or 10-memberedheterobicyclic group selected from: 1-azaspiro[3.3]heptan-1-yl,3-azabicyclo[3.1.0]hexan-3-yl, 3-azabicyclo[3.2.0]heptan-3-yl,3-azabicyclo[3.2.1]heptan-3-yl, 3-azabicyclo[3.2.1]octan-3-yl,3,4-dihydroisoquinolin-2(1H)-yl, 3,4-dihydro-2,7-naphthyridin-2(1H)-yl,5-azaspiro[2.4]heptan-5-yl, 5-azaspiro[2.5]octan-5-yl,6-azaspiro[2.5]octan-6-yl, 1-azaspiro[3.3]heptan-1-yl,6-azabicyclo[3.2.0]heptan-6-yl, isoindolin-2-yl, andoctahydrocyclopenta[c]pyrrol-2-yl, each of which is optionallysubstituted with one or two substituents independently selected from:C₁-C₃ alkoxy, C₁-C₃ alkyl, C₁-C₃ haloalkoxy, C₁-C₃ haloalkyl, andcarboxy.
 30. A compound according to claim 1, or a pharmaceuticallyacceptable salt, an optical isomer, or a mixture of optical isomersthereof, wherein R³ and R⁴, together with the nitrogen atom to whichthey are attached, form a 6, 7, 8, 9, or 10-membered heterobicyclicgroup selected from: 1-azaspiro[3.3]heptan-1-yl,3-azabicyclo[3.1.0]hexan-3-yl, 3-azabicyclo[3.2.0]heptan-3-yl,3-azabicyclo[3.2.1]heptan-3-yl, 3-azabicyclo[3.2.1]octan-3-yl,3,4-dihydroisoquinolin-2(1H)-yl, 3,4-dihydro-2,7-naphthyridin-2(1H)-yl,5-azaspiro[2.4]heptan-5-yl, 5-azaspiro[2.5]octan-5-yl,6-azaspiro[2.5]octan-6-yl, 1-azaspiro[3.3]heptan-1-yl,6-azabicyclo[3.2.0]heptan-6-yl, isoindolin-2-yl, andoctahydrocyclopenta[c]pyrrol-2-yl, each of which is optionallysubstituted with one or two substituents independently selected fromC₁-C₃ alkyl.
 31. A compound of formula:


32. A pharmaceutical composition comprising a compound according toclaim 1 or a pharmaceutically acceptable salt, an optical isomer, or amixture of optical isomers thereof and a pharmaceutically acceptablecarrier.
 33. A process for preparing a pharmaceutical compositioncomprising admixing a compound according to claim 1 or apharmaceutically acceptable salt, an optical isomer, or a mixture ofoptical isomers thereof and a pharmaceutically acceptable carrier.
 34. Amethod for treating a patient suffering from Huntington's disease,wherein the method comprises administering to the patient a compoundaccording to claim 1, or a pharmaceutically acceptable salt, an opticalisomer, or a mixture of optical isomers thereof.
 35. A method fortreating a patient suffering from Huntington's disease, wherein themethod comprises administering to the patient a compound according toclaim 4, or a pharmaceutically acceptable salt, an optical isomer, or amixture of optical isomers thereof.
 36. A method for treating a patientsuffering from Huntington's disease, wherein the method comprisesadministering to the patient a compound according to claim 31, or apharmaceutically acceptable salt, an optical isomer, or a mixture ofoptical isomers thereof.